TW202140551A

TW202140551A - Anti-CLDN18.2 antibody and use thereof used in the field of disease treatment and immunology for prevention and/or treatment of tumors

Info

Publication number: TW202140551A
Application number: TW109113197A
Authority: TW
Inventors: 杜靚; 張紅豔; 凌文; 袁紀軍
Original assignee: 大陸商上海吉倍生物技術有限公司
Priority date: 2020-04-20
Filing date: 2020-04-20
Publication date: 2021-11-01

Abstract

This invention relates to the field of disease treatment and immunology. Specifically, this invention relates to an anti-CLDN18.2 antibody or an antigen binding fragment thereof, a nucleic acid molecule for encoding the anti-CLDN18.2 antibody, an immunoconjugate, bispecific molecule, chimeric antigen receptor and pharmaceutical composition containing the anti-CLDN18.2 antibody as well as a use thereof for prevention and/or treatment of tumors.

Description

抗CLDN18.2抗體及其用途Anti-CLDN18.2 antibody and its use

本發明涉及疾病治療及免疫學領域，具體而言，本發明涉及抗CLDN18.2的抗體或其抗原結合片段，編碼其的核酸分子，包含其的免疫綴合物、雙特異性分子、嵌合抗原受體及藥物組合物，以及它們用於預防和/或治療腫瘤的用途。The present invention relates to the field of disease treatment and immunology. Specifically, the present invention relates to an anti-CLDN18. Antigen receptors and pharmaceutical compositions, and their use for the prevention and/or treatment of tumors.

胃癌是全球範圍內最常見的惡性腫瘤之一，其5年生存率低，絕大多數國家都在20%-40%的水準，每年約有70萬人死於該疾病。在中國胃癌的生存率為35.9%，據統計2015年中國胃癌的新發例數達679.1/10 萬，死亡例數高達498.0/10 萬，成為僅次於肺癌的高發病率及高死亡率的第二大腫瘤。胃癌的惡性程度較高，且由於胃癌篩查沒有廣泛開展，通常患者到晚期才得到診斷，錯失手術根治的機會，僅能以化療為主進行治療。Gastric cancer is one of the most common malignant tumors worldwide. Its 5-year survival rate is low. Most countries are at the level of 20%-40%. Approximately 700,000 people die from the disease every year. The survival rate of gastric cancer in China is 35.9%. According to statistics, the number of new cases of gastric cancer in China in 2015 reached 679.1/100,000, and the number of deaths was as high as 498.0/100,000, making it the highest incidence and death rate second only to lung cancer. The second largest tumor. The degree of malignancy of gastric cancer is relatively high, and because gastric cancer screening is not widely carried out, patients are usually diagnosed at an advanced stage, missing the opportunity for radical surgery, and can only be treated with chemotherapy.

隨著腫瘤分子生物學的發展，靶向藥物和免疫治療在血液瘤、乳腺癌、結直腸癌中的療效得以證實，但在胃癌治療中的應用相對緩慢。目前國際上已獲批胃癌靶向治療的靶點包括人類表皮生長因數受體2（HER2）和血管內皮生長因數（VEGF），獲批的胃癌免疫治療靶點為程式致死蛋白1（PD-1）。2017年9月，基於大型的三期臨床試驗ATTRACTION-2，日本厚生勞動省批准了PD-1抗體Opdivo用於治療化療耐藥的晚期胃癌患者：跟安慰劑相比，Opdivo可以降低37%的死亡率，有效率11.2%。基於二期臨床試驗Keynote 059，美國FDA加速批准了PD-1抗體Keytruda用於治療化療耐藥的PD-L1陽性的晚期胃癌患者：有效率15.5%。而在靶向治療方面，血管內皮生長因數（VEGF）不僅促進腫瘤血管產生，並且可以與腫瘤細胞表面的受體結合啟動下游訊號通路，直接參與腫瘤乾細胞的形成、發生與遷移等進程。VEGFR2抑制劑阿帕替尼和雷莫蘆單抗都已在臨床證明VEFR靶向治療在晚期胃癌二線及二線以上安全有效。胃癌每年的新發病例中約有6%-35% 出現HER2 基因擴增或蛋白過表達的現象，因此HER2是抗癌治療中重要的靶點之一。TOGA III期臨床試驗首次證實了曲妥珠單抗在HER2（+）晚期胃癌一線治療治療中的優勢，曲妥珠單抗聯合化療組的OS 較單純化療組明顯延長（13.8 個月vs.11 個月），次要終點包括PFS、ORR （客觀緩解率）、TTP（疾病進展時間）等也都有明顯改善。然而我國患者 HER2陽性的比例僅為10%左右，因此探索新的胃癌治療靶點勢在必行。With the development of tumor molecular biology, the efficacy of targeted drugs and immunotherapy in hematoma, breast cancer, and colorectal cancer has been confirmed, but the application in the treatment of gastric cancer has been relatively slow. At present, internationally approved targets for targeted therapy of gastric cancer include human epidermal growth factor receptor 2 (HER2) and vascular endothelial growth factor (VEGF). The approved immunotherapy targets for gastric cancer are programmed lethal protein 1 (PD-1). ). In September 2017, based on the large-scale phase III clinical trial ATTRACTION-2, the Ministry of Health, Labour and Welfare of Japan approved the PD-1 antibody Opdivo for the treatment of chemotherapy-resistant advanced gastric cancer patients: Compared with placebo, Opdivo can reduce 37% The mortality rate is 11.2%. Based on the Phase II clinical trial Keynote 059, the US FDA has accelerated the approval of the PD-1 antibody Keytruda for the treatment of chemotherapy-resistant PD-L1-positive advanced gastric cancer patients: the effective rate is 15.5%. In terms of targeted therapy, vascular endothelial growth factor (VEGF) not only promotes tumor blood vessel production, but also binds to receptors on the surface of tumor cells to initiate downstream signaling pathways, and directly participates in the formation, occurrence, and migration of tumor stem cells. Both VEGFR2 inhibitors apatinib and ramucirumab have been clinically proven that VEFR targeted therapy is safe and effective in the second-line and above-mentioned advanced gastric cancer. About 6%-35% of new cases of gastric cancer each year have HER2 gene amplification or protein overexpression, so HER2 is one of the important targets in anti-cancer therapy. The TOGA Phase III clinical trial confirmed the advantages of trastuzumab in the first-line treatment of HER2(+) advanced gastric cancer for the first time. The OS of the trastuzumab combined chemotherapy group was significantly longer than that of the chemotherapy alone group (13.8 months vs.11 Months), secondary endpoints including PFS, ORR (objective response rate), TTP (time to disease progression), etc. have also been significantly improved. However, the rate of HER2-positive patients in my country is only about 10%, so it is imperative to explore new therapeutic targets for gastric cancer.

密蛋白（Claudin）是上皮與內皮的緊密連接內的跨膜蛋白複合體，位於相鄰細胞間隙頂側，其分佈具有組織器官特異性，功能主要為細胞間黏附、維持細胞極性、調節細胞旁通透性及參與細胞增殖、分化的調節。密蛋白18（Claudin18）分子是四次跨膜蛋白，具有四個跨膜疏水區和兩個胞外環，以兩種不同剪接變體CLDN18.1和CLDN18.2存在。CLDN18.1和CLDN18.2在細胞內氮端和細胞外第一個胞外環存在序列上的差異，而其他部分的蛋白質一級序列相同。對CLDN18的組織分佈顯示，CLDN18.1僅在肺細胞中選擇性表達，而CLDN18.2僅在胃細胞上表達，並且CLDN18.2在正常胃中的表達局限於已分化的短期存在的胃上皮細胞中。CLDN18.2在胃細胞惡性轉化過程中經常性的被保留了，因而頻繁地在人胃癌細胞表面上被展示，60-80%的胃腸道腺瘤顯示CLDN18.2陽性（Clinical Cancer Research 2008, 14(23): 7624–34）。此外，CLDN18.2在胰管癌和轉移的胰腺癌上高表達，陽性率高達60-70%，可作為胰管/胰腺癌診斷標記和治療靶點（Journal of Clinical Pathology 2012, 65: 431-436; World Journal of Gastroenterology 2014, 20(31): 10813-10824; International Journal of Cancer 2014, 134: 731–739）。CLDN18.2在其他腫瘤上的異位元啟動尚無較為系統的研究，目前可查到的文獻顯示除上述的胰腺癌異位啟動這一蛋白在食道癌、支氣管癌、非小細胞肺癌（NSCLC)、乳腺癌、ENT腫瘤、卵巢癌、結腸癌、肝癌及其轉移癌，特別是胃癌轉移例如Krukenberg腫瘤、腹膜轉移和淋巴結轉移中均有發現表達（Clinical Cancer Research 2008, 14(23): 7624–34; International Journal of Cancer 2014, 134: 731–739; Cancer Letters 2017, 403: 66-73; International Journal of Cancer 2014, 135(9): 2206-2214）。CLDN18.2是具有預防和治療價值的腫瘤靶標，其在癌細胞和正常細胞之間的差異表達、其膜定位、其在大多數毒性相關正常組織中不存在、其在胃中的表達局限在可藉由胃的靶陰性乾細胞（或位置性不可及乾細胞）來補充的已分化的胃細胞，使得CLDN18.2成為癌症免疫治療的有吸引力的靶點。因此，發展具有更高的特異性、更低的毒副作用、更優的臨床藥效的抗CLDN18.2抗體是迫切而必要的，這將給癌症患者提供更多的用藥選擇。Claudin is a transmembrane protein complex in the tight junction of epithelium and endothelium. It is located on the top side of the adjacent intercellular space. Its distribution is specific to tissues and organs. Its function is mainly to adhere to cells, maintain cell polarity, and regulate cell side. Permeability and participate in the regulation of cell proliferation and differentiation. Claudin 18 (Claudin18) molecule is a four-pass transmembrane protein, with four transmembrane hydrophobic regions and two extracellular loops, and exists in two different splice variants CLDN18.1 and CLDN18.2. CLDN18.1 and CLDN18.2 have a sequence difference between the nitrogen end of the cell and the first extracellular loop outside the cell, while the protein primary sequence of the other parts is the same. The tissue distribution of CLDN18 shows that CLDN18.1 is only selectively expressed in lung cells, while CLDN18.2 is only expressed on gastric cells, and the expression of CLDN18.2 in normal stomachs is limited to differentiated short-term gastric epithelium. In the cell. CLDN18.2 is frequently retained during the malignant transformation of gastric cells, and is therefore frequently displayed on the surface of human gastric cancer cells. 60-80% of gastrointestinal adenomas are positive for CLDN18.2 (Clinical Cancer Research 2008, 14 (23): 7624–34). In addition, CLDN18.2 is highly expressed on pancreatic duct cancer and metastatic pancreatic cancer, with a positive rate of 60-70%, which can be used as a diagnostic marker and therapeutic target for pancreatic duct/pancreatic cancer (Journal of Clinical Pathology 2012, 65: 431- 436; World Journal of Gastroenterology 2014, 20(31): 10813-10824; International Journal of Cancer 2014, 134: 731–739). There is no systematic study on the ectopic initiation of CLDN18.2 on other tumors. The currently available literature shows that in addition to the above-mentioned pancreatic cancer ectopic initiation, this protein is involved in esophageal cancer, bronchial cancer, and non-small cell lung cancer (NSCLC). ), breast cancer, ENT tumor, ovarian cancer, colon cancer, liver cancer and its metastases, especially gastric cancer metastasis such as Krukenberg tumor, peritoneal metastasis and lymph node metastasis have been found to express (Clinical Cancer Research 2008, 14(23): 7624 –34; International Journal of Cancer 2014, 134: 731–739; Cancer Letters 2017, 403: 66-73; International Journal of Cancer 2014, 135(9): 2206-2214). CLDN 18.2 is a tumor target with preventive and therapeutic value. Its differential expression between cancer cells and normal cells, its membrane location, its absence in most normal tissues related to toxicity, and its expression in the stomach are limited Differentiated gastric cells, which can be supplemented by target-negative stem cells (or positionally inaccessible stem cells) of the stomach, make CLDN 18.2 an attractive target for cancer immunotherapy. Therefore, it is urgent and necessary to develop anti-CLDN18.2 antibodies with higher specificity, lower side effects, and better clinical efficacy, which will provide cancer patients with more medication options.

本發明的抗體能夠特異性識別/結合人CLDN18.2，並且能夠藉由ADCC和/或CDC來誘導殺傷表達CLDN18.2的細胞（例如，腫瘤細胞），且與已知的抗CLDN18.2抗體相比具有更優的功能特性。因此，本發明的抗體具有用於預防和/或治療腫瘤的潛力，具有重大的臨床價值。本發明的抗體The antibody of the present invention can specifically recognize/bind human CLDN18.2, and can induce and kill CLDN18.2-expressing cells (for example, tumor cells) by ADCC and/or CDC, and is compatible with known anti-CLDN18.2 antibodies Compared to have better functional characteristics. Therefore, the antibody of the present invention has the potential for preventing and/or treating tumors, and has great clinical value. Antibodies of the invention

因此，在一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 3，或與SEQ ID NO: 3相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 4，或與SEQ ID NO: 4相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 5，或與SEQ ID NO: 5相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 6，或與SEQ ID NO: 6相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 7，或與SEQ ID NO: 7相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 8，或與SEQ ID NO: 8相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。Therefore, in one aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 3, or with one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3) compared with SEQ ID NO: 3 Amino acid substitution, deletion or addition) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 4, or with one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3) compared with SEQ ID NO: 4 Amino acid substitution, deletion or addition) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 5, or with one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3) compared with SEQ ID NO: 5 Amino acid substitution, deletion or addition) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 6, or compared with SEQ ID NO: 6 with one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 Amino acid substitution, deletion or addition) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 7, or compared with SEQ ID NO: 7 with one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 Amino acid substitution, deletion or addition) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 8, or compared with SEQ ID NO: 8 with one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 Amino acid substitution, deletion or addition) sequence.

在某些較佳的實施方案中，(i)-(vi)任一項中所述的置換為保守置換；在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 3所示的VH CDR1、如SEQ ID NO: 4所示的VH CDR2、如SEQ ID NO: 5所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 6或96所示的VL CDR1、如SEQ ID NO: 7所示的VL CDR2、如SEQ ID NO: 8所示的VL CDR3。In some preferred embodiments, the substitutions described in any one of (i)-(vi) are conservative substitutions; In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 shown in SEQ ID NO: 3, the VH CDR2 shown in SEQ ID NO: 4, and the VH CDR2 shown in SEQ ID NO: VH CDR3 shown in 5; and, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 6 or 96, VL CDR2 as shown in SEQ ID NO: 7 and VL CDR2 as shown in SEQ ID NO : VL CDR3 shown in 8.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中， (a) 該重鏈可變區包含SEQ ID NO: 1所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 2所示的輕鏈可變區中含有的3個CDR；或 (b) 該重鏈可變區包含SEQ ID NO: 91所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 92所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein, (a) The heavy chain variable region includes the 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 1; and, the light chain variable region includes the light chain variable region shown in SEQ ID NO: 2 3 CDRs contained in the variable region; or (b) The heavy chain variable region includes the 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 91; and, the light chain variable region includes the light chain variable region shown in SEQ ID NO: 92 The 3 CDRs contained in the variable region.

在某些較佳的實施方案中，該重鏈可變區中含有的3個CDR，和/或該輕鏈可變區中含有的3個CDR，由Kabat、Chothia或IMGT編號系統定義。In some preferred embodiments, the 3 CDRs contained in the heavy chain variable region and/or the 3 CDRs contained in the light chain variable region are defined by the Kabat, Chothia or IMGT numbering system.

在某些較佳的實施方案中，該抗體或其抗原結合片段是44F7或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 44F7 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段具備以下生物學功能中的一項或多項： (a) 以0.1 μg/ml或更小（例如0.05 μg/ml或更小）的EC50結合人CLDN18.2； (b) 以0.1 μg/ml或更小的EC50結合小鼠CLDN18.2； (c) 不結合人CLDN18.1； (d) 藉由抗體依賴性細胞介導的細胞毒性（ADCC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (e) 藉由補體依賴的細胞毒性（CDC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (f) 在受試者中預防和/治療腫瘤（例如，表達CLDN18.2的腫瘤）。In some preferred embodiments, the antibody or antigen-binding fragment thereof has one or more of the following biological functions: (a) Binding to human CLDN 18.2 with an EC50 of 0.1 μg/ml or less (for example, 0.05 μg/ml or less); (b) Binding mouse CLDN18.2 with an EC50 of 0.1 μg/ml or less; (c) Does not combine with human CLDN 18.1; (d) Antibody-dependent cell-mediated cytotoxicity (ADCC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (e) Complement-dependent cytotoxicity (CDC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (f) Preventing and/treating tumors in subjects (for example, tumors expressing CLDN 18.2).

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 3所示的VH CDR1、如SEQ ID NO: 4所示的VH CDR2、如SEQ ID NO: 5所示的VH CDR3；如SEQ ID NO: 6或96所示的VL CDR1、如SEQ ID NO: 7所示的VL CDR2、如SEQ ID NO: 8所示的VL CDR3；或 (2) (a)SEQ ID NO: 1所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 2所示的輕鏈可變區中含有的3個CDR；或(b) SEQ ID NO: 91所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 92所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 as shown in SEQ ID NO: 3, VH CDR2 as shown in SEQ ID NO: 4, VH CDR3 as shown in SEQ ID NO: 5; as shown in SEQ ID NO: 6 or 96 VL CDR1, VL CDR2 as shown in SEQ ID NO: 7 and VL CDR3 as shown in SEQ ID NO: 8; or (2) (a) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 1; and, the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 2; or ( b) 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 91; and 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 92.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 1所示的序列； (ii) 與SEQ ID NO: 1所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 1所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 2所示的序列； (v) 與SEQ ID NO: 2所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 2所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 1; (ii) Compared with the sequence shown in SEQ ID NO: 1, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 1 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 2; (v) Compared with the sequence shown in SEQ ID NO: 2, there are one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 2 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，(ii)或(v)中所述的置換是保守置換。In certain preferred embodiments, the substitutions described in (ii) or (v) are conservative substitutions.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 1所示的序列的VH和具有如SEQ ID NO:2所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 1 and VL having the sequence shown in SEQ ID NO: 2.

在某些較佳的實施方案中，該抗體或其抗原結合片段是人源化的。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is humanized.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含來源於人免疫球蛋白的重鏈可變區（VH）構架區（FR），和/或該抗體或其抗原結合片段的VL包含來源於人免疫球蛋白的輕鏈可變區（VL）構架區（FR）。在此類實施方案中，該抗體或其抗原結合片段的重鏈可變區FR和/或輕鏈可變區FR可以包含一或複數非人源（例如，鼠源）氨基酸殘基，例如該重鏈構架區FR和/或輕鏈構架區FR可以包含一或複數氨基酸回復突變，在這些回復突變中有相應的鼠源氨基酸殘基。In certain preferred embodiments, the VH of the antibody or its antigen-binding fragment comprises a heavy chain variable region (VH) framework region (FR) derived from human immunoglobulin, and/or the antibody or its antigen-binding The VL of the fragment contains a light chain variable region (VL) framework region (FR) derived from human immunoglobulin. In such embodiments, the heavy chain variable region FR and/or light chain variable region FR of the antibody or antigen-binding fragment thereof may comprise one or more non-human (e.g., murine) amino acid residues, such as the The FR of the heavy chain framework region and/or the FR of the light chain framework region may contain one or more amino acid back mutations, in which there are corresponding murine amino acid residues.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (a) 人免疫球蛋白的重鏈構架區或其變體，該變體與其所源自的序列相比具有至多20個氨基酸的保守置換（例如至多15個、至多10個、或至多5個氨基酸的保守置換；例如1個、2個、3個、4個或5個氨基酸的保守置換）；和/或 (b) 人免疫球蛋白的輕鏈構架區或其變體，該變體與其所源自的序列相比具有至多20個氨基酸的保守置換（例如至多15個、至多10個、或至多5個氨基酸的保守置換；例如1個、2個、3個、4個或5個氨基酸的保守置換）。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (a) Human immunoglobulin heavy chain framework regions or variants thereof, which have conservative substitutions of up to 20 amino acids compared to the sequence from which they are derived (for example, up to 15, up to 10, or up to 5 Conservative substitutions of amino acids; for example, conservative substitutions of 1, 2, 3, 4, or 5 amino acids); and/or (b) The light chain framework region of a human immunoglobulin or a variant thereof, which has conservative substitutions of up to 20 amino acids compared to the sequence from which it is derived (for example, up to 15, up to 10, or up to 5 Conservative substitutions of amino acids; for example, conservative substitutions of 1, 2, 3, 4, or 5 amino acids).

在某些較佳的實施方案中，該抗體或其抗原結合片段包含人免疫球蛋白的構架區，例如人胚系抗體基因所編碼的氨基酸序列中所包含的構架區。在某些較佳的實施方案中，該抗體或其抗原結合片段包含：人重鏈胚系基因所編碼的氨基酸序列中所包含的重鏈構架區，和/或人輕鏈胚系基因所編碼的氨基酸序列中所包含的輕鏈構架區。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises a framework region of a human immunoglobulin, for example, a framework region contained in an amino acid sequence encoded by a human germline antibody gene. In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: the heavy chain framework region contained in the amino acid sequence encoded by the human heavy chain germline gene, and/or the human light chain germline gene encoded The light chain framework region contained in the amino acid sequence of.

在此類實施方案中，該抗體或其抗原結合片段的構架區（重鏈構架區和/或輕鏈構架區）可以包含一或複數非人源（例如，鼠源）氨基酸殘基。在某些較佳的實施方案中，該構架區（重鏈構架區和/或輕鏈構架區）包含一或複數氨基酸殘基，其被回復突變至相應的鼠源殘基或相應鼠源殘基的保守氨基酸置換（此類突變稱為回復突變（back mutation））。In such embodiments, the framework regions (heavy chain framework regions and/or light chain framework regions) of the antibody or antigen-binding fragment thereof may comprise one or more non-human (eg, murine) amino acid residues. In some preferred embodiments, the framework region (heavy chain framework region and/or light chain framework region) contains one or more amino acid residues, which are backmutated to corresponding murine residues or corresponding murine residues Conservative amino acid substitutions (such mutations are called back mutations).

因此，在某些較佳的實施方案中，該抗體或其抗原結合片段包含人免疫球蛋白的構架區（例如，人胚系抗體基因所編碼的氨基酸序列中所包含的構架區），所述構架區任選地包含一或複數從人源殘基至鼠源殘基的回復突變。Therefore, in certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises a framework region of human immunoglobulin (for example, a framework region contained in an amino acid sequence encoded by a human germline antibody gene), said The framework region optionally contains one or more back mutations from human residues to murine residues.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 91所示的序列； (ii) 與SEQ ID NO: 91所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 91所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 92所示的序列； (v) 與SEQ ID NO: 92所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 92所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 91; (ii) Compared with the sequence shown in SEQ ID NO: 91, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 91 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 92; (v) Compared with the sequence shown in SEQ ID NO: 92, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 92 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 91所示的序列的VH和具有如SEQ ID NO: 92所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 91 and VL having the sequence shown in SEQ ID NO: 92.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 99所示的序列； (ii) 與SEQ ID NO: 99所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 99所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 100所示的序列； (v) 與SEQ ID NO: 100所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 100所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 99; (ii) Compared with the sequence shown in SEQ ID NO: 99, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 99 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 100; (v) Compared with the sequence shown in SEQ ID NO: 100, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 100 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 99所示的序列的VH和具有如SEQ ID NO: 100所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 99 and VL having the sequence shown in SEQ ID NO: 100.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 101所示的序列； (ii) 與SEQ ID NO: 101所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 101所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 102所示的序列； (v) 與SEQ ID NO: 102所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 102所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 101; (ii) Compared with the sequence shown in SEQ ID NO: 101, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 101 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 102; (v) Compared with the sequence shown in SEQ ID NO: 102, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 102 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 101所示的序列的VH和具有如SEQ ID NO: 102所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 101 and VL having the sequence shown in SEQ ID NO: 102.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 75，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 76，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 77，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 78，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 79，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 80，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 75, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 76, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 77, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 78, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 79, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 80, or has one or several amino acid substitutions, deletions, or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，(i)-(vi)任一項中所述的置換為保守置換。In certain preferred embodiments, the substitutions described in any one of (i)-(vi) are conservative substitutions.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 75所示的VH CDR1、如SEQ ID NO: 76所示的VH CDR2、如SEQ ID NO: 77所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 78所示的VL CDR1、如SEQ ID NO: 79所示的VL CDR2、如SEQ ID NO: 80所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 75, the VH CDR2 as shown in SEQ ID NO: 76, and the VH CDR2 as shown in SEQ ID NO: The VH CDR3 shown in 77; and, the VL of the antibody or antigen-binding fragment thereof includes: VL CDR1 shown in SEQ ID NO: 78, VL CDR2 shown in SEQ ID NO: 79, and VL CDR2 shown in SEQ ID NO: 80 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 73所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 74所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 73; and the light chain variable region includes 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 74.

在某些較佳的實施方案中，該抗體或其抗原結合片段是1D10或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 1D10 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段具備以下生物學功能中的一項或多項： (a) 以0.1 μg/ml或更小（例如，0.05 μg/ml、0.02 μg/ml或更小）的EC50結合人CLDN18.2； (b) 以1 μg/ml或更小的EC50結合小鼠CLDN18.2； (c) 藉由抗體依賴性細胞介導的細胞毒性（ADCC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (d) 藉由補體依賴的細胞毒性（CDC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (e) 介導CLDN18.2內化至細胞（例如，腫瘤細胞）中，例如以如藉由FACS或流式細胞術所測量的至少10%（例如至少15%，至少20%或更多）的內化水準；該細胞在其表面表達CLDN18.2； (f) 在受試者中預防和/治療腫瘤（例如，表達CLDN18.2的腫瘤）。In some preferred embodiments, the antibody or antigen-binding fragment thereof has one or more of the following biological functions: (a) Binding to human CLDN 18.2 with an EC50 of 0.1 μg/ml or less (for example, 0.05 μg/ml, 0.02 μg/ml or less); (b) Binding mouse CLDN18.2 with an EC50 of 1 μg/ml or less; (c) Using antibody-dependent cell-mediated cytotoxicity (ADCC) to induce and kill cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (d) Complement-dependent cytotoxicity (CDC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (e) Mediate the internalization of CLDN 18.2 into cells (e.g., tumor cells), for example at least 10% (e.g. at least 15%, at least 20% or more) as measured by FACS or flow cytometry The internalization level of the cell; the cell expresses CLDN18.2 on its surface; (f) Preventing and/treating tumors in subjects (for example, tumors expressing CLDN 18.2).

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 75所示的VH CDR1、如SEQ ID NO: 76所示的VH CDR2、如SEQ ID NO: 77所示的VH CDR3；如SEQ ID NO: 78所示的VL CDR1、如SEQ ID NO: 79所示的VL CDR2、如SEQ ID NO: 80所示的VL CDR3；或 (2) SEQ ID NO: 73所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 74所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 75, VH CDR2 shown in SEQ ID NO: 76, VH CDR3 shown in SEQ ID NO: 77; VL CDR1 shown in SEQ ID NO: 78 , VL CDR2 as shown in SEQ ID NO: 79, VL CDR3 as shown in SEQ ID NO: 80; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 73; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 74.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 73所示的序列； (ii) 與SEQ ID NO: 73所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 73所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 74所示的序列； (v) 與SEQ ID NO: 74所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 74所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 73; (ii) Compared with the sequence shown in SEQ ID NO: 73, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 73 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 74; (v) Compared with the sequence shown in SEQ ID NO: 74, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 74 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 73所示的序列的VH和具有如SEQ ID NO: 74所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 73 and VL having the sequence shown in SEQ ID NO: 74.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 11，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 12，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 13，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 14，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 15，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 16，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 11, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 12, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 13, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 14, or compared with it, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 15, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 16, or compared with SEQ ID NO: 16, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 11所示的VH CDR1、如SEQ ID NO: 12所示的VH CDR2、如SEQ ID NO: 13所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 14所示的VL CDR1、如SEQ ID NO: 15所示的VL CDR2、如SEQ ID NO: 16所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 11, the VH CDR2 as shown in SEQ ID NO: 12, and the VH CDR2 as shown in SEQ ID NO: The VH CDR3 shown in 13; and, the VL of the antibody or the antigen-binding fragment thereof includes: VL CDR1 shown in SEQ ID NO: 14, VL CDR2 shown in SEQ ID NO: 15, and VL CDR2 shown in SEQ ID NO: 16 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 9所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 10所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 9; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 10.

在某些較佳的實施方案中，該抗體或其抗原結合片段是2F12或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 2F12 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段具備以下生物學功能中的一項或多項： (a) 以0.1 μg/ml或更小（例如，0.05 μg/ml或更小）的EC50結合人CLDN18.2； (b) 以0.1 μg/ml或更小的EC50結合小鼠CLDN18.2； (c) 不結合人CLDN18.1； (d) 藉由抗體依賴性細胞介導的細胞毒性（ADCC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (e) 藉由補體依賴的細胞毒性（CDC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (f) 在受試者中預防和/治療腫瘤（例如，表達CLDN18.2的腫瘤）。In some preferred embodiments, the antibody or antigen-binding fragment thereof has one or more of the following biological functions: (a) Binding to human CLDN 18.2 with an EC50 of 0.1 μg/ml or less (for example, 0.05 μg/ml or less); (b) Binding mouse CLDN18.2 with an EC50 of 0.1 μg/ml or less; (c) Does not combine with human CLDN 18.1; (d) Antibody-dependent cell-mediated cytotoxicity (ADCC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (e) Complement-dependent cytotoxicity (CDC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (f) Preventing and/treating tumors in subjects (for example, tumors expressing CLDN 18.2).

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 11所示的VH CDR1、如SEQ ID NO: 12所示的VH CDR2、如SEQ ID NO: 13所示的VH CDR3；如SEQ ID NO: 14所示的VL CDR1、如SEQ ID NO: 15所示的VL CDR2、如SEQ ID NO: 16所示的VL CDR3；或 (2) SEQ ID NO: 9所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 10所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 11, VH CDR2 shown in SEQ ID NO: 12, VH CDR3 shown in SEQ ID NO: 13; VL CDR1 shown in SEQ ID NO: 14 , VL CDR2 as shown in SEQ ID NO: 15 and VL CDR3 as shown in SEQ ID NO: 16; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 9; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 10.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 9所示的序列； (ii) 與SEQ ID NO: 9所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 9所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 10所示的序列； (v) 與SEQ ID NO: 10所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 10所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 9; (ii) Compared with the sequence shown in SEQ ID NO: 9 has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions ) Sequence; or (iii) The sequence shown in SEQ ID NO: 9 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 10; (v) Compared with the sequence shown in SEQ ID NO: 10, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 10 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 9所示的序列的VH和具有如SEQ ID NO: 10所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 9 and VL having the sequence shown in SEQ ID NO: 10.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 19，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 20，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 21，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 22，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 23，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 24，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 19, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 20, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 21, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 22, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 23, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 24, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 19所示的VH CDR1、如SEQ ID NO: 20所示的VH CDR2、如SEQ ID NO: 21所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 22所示的VL CDR1、如SEQ ID NO: 23所示的VL CDR2、如SEQ ID NO: 24所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 19, the VH CDR2 as shown in SEQ ID NO: 20, and the VH CDR2 as shown in SEQ ID NO: VH CDR3 shown in 21; and, the VL of the antibody or antigen-binding fragment thereof includes: VL CDR1 as shown in SEQ ID NO: 22, VL CDR2 as shown in SEQ ID NO: 23, and VL CDR2 as shown in SEQ ID NO: 24 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 17所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 18所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 17; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 18.

在某些較佳的實施方案中，該抗體或其抗原結合片段是3F2或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 3F2 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 19所示的VH CDR1、如SEQ ID NO: 20所示的VH CDR2、如SEQ ID NO: 21所示的VH CDR3；如SEQ ID NO: 22所示的VL CDR1、如SEQ ID NO: 23所示的VL CDR2、如SEQ ID NO: 24所示的VL CDR3；或 (2) SEQ ID NO: 17所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 18所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 19, VH CDR2 shown in SEQ ID NO: 20, VH CDR3 shown in SEQ ID NO: 21; VL CDR1 shown in SEQ ID NO: 22 , VL CDR2 as shown in SEQ ID NO: 23, VL CDR3 as shown in SEQ ID NO: 24; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 17; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 18.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 17所示的序列； (ii) 與SEQ ID NO: 17所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 17所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 18所示的序列； (v) 與SEQ ID NO: 18所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 18所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 17; (ii) Compared with the sequence shown in SEQ ID NO: 17 has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions ) Sequence; or (iii) The sequence shown in SEQ ID NO: 17 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 18; (v) Compared with the sequence shown in SEQ ID NO: 18, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 18 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 17所示的序列的VH和具有如SEQ ID NO: 18所示的序列的VL。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 17 and VL having the sequence shown in SEQ ID NO: 18.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 27，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 28，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 29，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 30，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 31，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 32，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 27, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 28, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 29, or has one or several amino acid substitutions, deletions, or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 30, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 31, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 32, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 27所示的VH CDR1、如SEQ ID NO: 28所示的VH CDR2、如SEQ ID NO: 29所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 30所示的VL CDR1、如SEQ ID NO: 31所示的VL CDR2、如SEQ ID NO: 32所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 27, the VH CDR2 as shown in SEQ ID NO: 28, and the VH CDR2 as shown in SEQ ID NO: The VH CDR3 shown in 29; and the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 shown in SEQ ID NO: 30, VL CDR2 shown in SEQ ID NO: 31, and VL CDR2 shown in SEQ ID NO: 32 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 25所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 26所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 25; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 26.

在某些較佳的實施方案中，該抗體或其抗原結合片段是5F9或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 5F9 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段具備以下生物學功能中的一項或多項： (a) 以0.5 μg/ml或更小的EC50結合人CLDN18.2； (b) 以0.2 μg/ml或更小的EC50結合小鼠CLDN18.2； (c) 藉由抗體依賴性細胞介導的細胞毒性（ADCC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (d) 藉由補體依賴的細胞毒性（CDC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (e) 介導CLDN18.2內化至細胞（例如，腫瘤細胞）中，例如以如藉由FACS或流式細胞術所測量的至少10%（例如至少15%，至少20%或更多）的內化水準；該細胞在其表面表達CLDN18.2； (f) 在受試者中預防和/治療腫瘤（例如，表達CLDN18.2的腫瘤）。In some preferred embodiments, the antibody or antigen-binding fragment thereof has one or more of the following biological functions: (a) Binding to human CLDN 18.2 with an EC50 of 0.5 μg/ml or less; (b) Binding mouse CLDN18.2 with an EC50 of 0.2 μg/ml or less; (c) Using antibody-dependent cell-mediated cytotoxicity (ADCC) to induce and kill cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (d) Complement-dependent cytotoxicity (CDC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (e) Mediate the internalization of CLDN 18.2 into cells (e.g., tumor cells), for example at least 10% (e.g. at least 15%, at least 20% or more) as measured by FACS or flow cytometry The internalization level of the cell; the cell expresses CLDN18.2 on its surface; (f) Preventing and/treating tumors in subjects (for example, tumors expressing CLDN 18.2).

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 27所示的VH CDR1、如SEQ ID NO: 28所示的VH CDR2、如SEQ ID NO: 29所示的VH CDR3；如SEQ ID NO: 30所示的VL CDR1、如SEQ ID NO: 31所示的VL CDR2、如SEQ ID NO: 32所示的VL CDR3；或 (2) SEQ ID NO: 25所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 26所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 27, VH CDR2 shown in SEQ ID NO: 28, VH CDR3 shown in SEQ ID NO: 29; VL CDR1 shown in SEQ ID NO: 30 , VL CDR2 as shown in SEQ ID NO: 31, VL CDR3 as shown in SEQ ID NO: 32; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 25; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 26.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 25所示的序列； (ii) 與SEQ ID NO: 25所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 25所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 26所示的序列； (v) 與SEQ ID NO: 26所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 26所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 25; (ii) Compared with the sequence shown in SEQ ID NO: 25, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 25 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 26; (v) Compared with the sequence shown in SEQ ID NO: 26, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 26 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 25所示的序列的VH和具有如SEQ ID NO: 26所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 25 and VL having the sequence shown in SEQ ID NO: 26.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 35，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 36，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 37，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 38，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 39，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 40，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 35, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 36, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 37, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 38, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 39, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 40, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 35所示的VH CDR1、如SEQ ID NO: 36所示的VH CDR2、如SEQ ID NO: 37所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 38所示的VL CDR1、如SEQ ID NO: 39所示的VL CDR2、如SEQ ID NO: 40所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 35, the VH CDR2 as shown in SEQ ID NO: 36, and the VH CDR2 as shown in SEQ ID NO: The VH CDR3 shown in 37; and, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 shown in SEQ ID NO: 38, VL CDR2 shown in SEQ ID NO: 39, and VL CDR2 shown in SEQ ID NO: 40 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 33所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 34所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 33; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 34.

在某些較佳的實施方案中，該抗體或其抗原結合片段是9F3或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 9F3 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段具備以下生物學功能中的一項或多項： (a) 以0.2 μg/ml或更小的EC50結合人CLDN18.2； (b) 以0.1 μg/ml或更小的EC50結合小鼠CLDN18.2； (c) 不結合人CLDN18.1； (d) 藉由抗體依賴性細胞介導的細胞毒性（ADCC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (e) 藉由補體依賴的細胞毒性（CDC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (f) 在受試者中預防和/治療腫瘤（例如，表達CLDN18.2的腫瘤）。In some preferred embodiments, the antibody or antigen-binding fragment thereof has one or more of the following biological functions: (a) Binding to human CLDN 18.2 with an EC50 of 0.2 μg/ml or less; (b) Binding mouse CLDN18.2 with an EC50 of 0.1 μg/ml or less; (c) Does not combine with human CLDN 18.1; (d) Antibody-dependent cell-mediated cytotoxicity (ADCC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (e) Complement-dependent cytotoxicity (CDC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (f) Preventing and/treating tumors in subjects (for example, tumors expressing CLDN 18.2).

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 35所示的VH CDR1、如SEQ ID NO: 36所示的VH CDR2、如SEQ ID NO: 37所示的VH CDR3；如SEQ ID NO: 38所示的VL CDR1、如SEQ ID NO: 39所示的VL CDR2、如SEQ ID NO: 40所示的VL CDR3；或 (2) SEQ ID NO: 33所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 34所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 35, VH CDR2 shown in SEQ ID NO: 36, VH CDR3 shown in SEQ ID NO: 37; VL CDR1 shown in SEQ ID NO: 38 , VL CDR2 as shown in SEQ ID NO: 39, VL CDR3 as shown in SEQ ID NO: 40; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 33; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 34.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 33所示的序列； (ii) 與SEQ ID NO: 33所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 33所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 34所示的序列； (v) 與SEQ ID NO: 34所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 34所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 33; (ii) Compared with the sequence shown in SEQ ID NO: 33, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 33 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 34; (v) Compared with the sequence shown in SEQ ID NO: 34, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 34 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 33所示的序列的VH和具有如SEQ ID NO: 34所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 33 and VL having the sequence shown in SEQ ID NO: 34.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 43，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 44，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 45，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 46，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 47，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 48，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 43, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 44, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 45, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 46, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 47, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 48, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 43所示的VH CDR1、如SEQ ID NO: 44所示的VH CDR2、如SEQ ID NO: 45所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 46所示的VL CDR1、如SEQ ID NO: 47所示的VL CDR2、如SEQ ID NO: 48所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 shown in SEQ ID NO: 43, the VH CDR2 shown in SEQ ID NO: 44, and the VH CDR2 shown in SEQ ID NO: The VH CDR3 shown in 45; and, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 shown in SEQ ID NO: 46, VL CDR2 shown in SEQ ID NO: 47, and VL CDR2 shown in SEQ ID NO: 48 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 41所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 42所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 41; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 42.

在某些較佳的實施方案中，該抗體或其抗原結合片段是10B11或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 10B11 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 43所示的VH CDR1、如SEQ ID NO: 44所示的VH CDR2、如SEQ ID NO: 45所示的VH CDR3；如SEQ ID NO: 46所示的VL CDR1、如SEQ ID NO: 47所示的VL CDR2、如SEQ ID NO: 48所示的VL CDR3；或 (2) SEQ ID NO: 41所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 42所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 43, VH CDR2 shown in SEQ ID NO: 44, VH CDR3 shown in SEQ ID NO: 45; VL CDR1 shown in SEQ ID NO: 46 , VL CDR2 as shown in SEQ ID NO: 47, VL CDR3 as shown in SEQ ID NO: 48; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 41; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 42.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 41所示的序列； (ii) 與SEQ ID NO: 41所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 41所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 42所示的序列； (v) 與SEQ ID NO: 42所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 42所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 41; (ii) Compared with the sequence shown in SEQ ID NO: 41, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 41 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 42; (v) Compared with the sequence shown in SEQ ID NO: 42 there are one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions ) Sequence; or (vi) The sequence shown in SEQ ID NO: 42 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 41所示的序列的VH和具有如SEQ ID NO: 42所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 41 and VL having the sequence shown in SEQ ID NO: 42.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 51，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 52，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 53，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 54，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 55，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 56，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 51, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 52, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 53, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 54, or compared with it, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 55, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 56, or compared with it, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 51所示的VH CDR1、如SEQ ID NO: 52所示的VH CDR2、如SEQ ID NO: 53所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 54所示的VL CDR1、如SEQ ID NO: 55所示的VL CDR2、如SEQ ID NO: 56所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 shown in SEQ ID NO: 51, the VH CDR2 shown in SEQ ID NO: 52, and the VH CDR2 shown in SEQ ID NO: The VH CDR3 shown in 53; and, the VL of the antibody or the antigen-binding fragment thereof includes: VL CDR1 shown in SEQ ID NO: 54; VL CDR2 shown in SEQ ID NO: 55; VL CDR2 shown in SEQ ID NO: 56 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 49所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 50所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 49; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 50.

在某些較佳的實施方案中，該抗體或其抗原結合片段是27B5或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 27B5 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 51所示的VH CDR1、如SEQ ID NO: 52所示的VH CDR2、如SEQ ID NO: 53所示的VH CDR3；如SEQ ID NO: 54所示的VL CDR1、如SEQ ID NO: 55所示的VL CDR2、如SEQ ID NO: 56所示的VL CDR3；或 (2) SEQ ID NO: 49所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 50所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 51, VH CDR2 shown in SEQ ID NO: 52, VH CDR3 shown in SEQ ID NO: 53; VL CDR1 shown in SEQ ID NO: 54 , VL CDR2 as shown in SEQ ID NO: 55, VL CDR3 as shown in SEQ ID NO: 56; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 49; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 50.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 49所示的序列； (ii) 與SEQ ID NO: 49所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 49所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 50所示的序列； (v) 與SEQ ID NO: 50所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 50所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 49; (ii) Compared with the sequence shown in SEQ ID NO: 49, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 49 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 50; (v) Compared with the sequence shown in SEQ ID NO: 50, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 50 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 49所示的序列的VH和具有如SEQ ID NO: 50所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 49 and VL having the sequence shown in SEQ ID NO: 50.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 59，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 60，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 61，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 62，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 63，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 64，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 59, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 60, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 61, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 62, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 63, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 64, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 59所示的VH CDR1、如SEQ ID NO: 60所示的VH CDR2、如SEQ ID NO: 61所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 62所示的VL CDR1、如SEQ ID NO: 63所示的VL CDR2、如SEQ ID NO: 64所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 shown in SEQ ID NO: 59, the VH CDR2 shown in SEQ ID NO: 60, and the VH CDR2 shown in SEQ ID NO: The VH CDR3 shown in 61; and, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 shown in SEQ ID NO: 62, VL CDR2 shown in SEQ ID NO: 63, and VL CDR2 shown in SEQ ID NO: 64 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 57所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 58所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The three CDRs contained in the heavy chain variable region shown in SEQ ID NO: 57; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 58.

在某些較佳的實施方案中，該抗體或其抗原結合片段是37B1或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In certain preferred embodiments, the antibody or antigen-binding fragment thereof is 37B1 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 59所示的VH CDR1、如SEQ ID NO: 60所示的VH CDR2、如SEQ ID NO: 61所示的VH CDR3；如SEQ ID NO: 62所示的VL CDR1、如SEQ ID NO: 63所示的VL CDR2、如SEQ ID NO: 64所示的VL CDR3；或 (2) SEQ ID NO: 57所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 58所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 59, VH CDR2 shown in SEQ ID NO: 60, VH CDR3 shown in SEQ ID NO: 61; VL CDR1 shown in SEQ ID NO: 62 , VL CDR2 as shown in SEQ ID NO: 63, VL CDR3 as shown in SEQ ID NO: 64; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 57; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 58.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 57所示的序列； (ii) 與SEQ ID NO: 57所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 57所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 58所示的序列； (v) 與SEQ ID NO: 58所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 58所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 57; (ii) Compared with the sequence shown in SEQ ID NO: 57, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 57 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 58; (v) Compared with the sequence shown in SEQ ID NO: 58 there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 58 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 57所示的序列的VH和具有如SEQ ID NO: 58所示的序列的VL。In some preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 57 and VL having the sequence shown in SEQ ID NO: 58.

在另一個方面，本發明提供了能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 67，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 68，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 69，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 70，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 71，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 72，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列。In another aspect, the present invention provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN 18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 67, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 68, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 69, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 70, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 71, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 72, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence.

在某些較佳的實施方案中，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 67所示的VH CDR1、如SEQ ID NO: 68所示的VH CDR2、如SEQ ID NO: 69所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 70所示的VL CDR1、如SEQ ID NO: 71所示的VL CDR2、如SEQ ID NO: 72所示的VL CDR3。In some preferred embodiments, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 67, the VH CDR2 as shown in SEQ ID NO: 68, and the VH CDR2 as shown in SEQ ID NO: The VH CDR3 shown in 69; and, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 shown in SEQ ID NO: 70, VL CDR2 shown in SEQ ID NO: 71, and VL CDR2 shown in SEQ ID NO: 72 VL CDR3 shown.

本發明還提供了一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含重鏈可變區和輕鏈可變區，其中，該重鏈可變區包含SEQ ID NO: 65所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 66所示的輕鏈可變區中含有的3個CDR。The present invention also provides an antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2. The antibody or antigen-binding fragment thereof comprises a heavy chain variable region and a light chain variable region, wherein the heavy chain variable region comprises The three CDRs contained in the heavy chain variable region shown in SEQ ID NO: 65; and the light chain variable region includes the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 66.

在某些較佳的實施方案中，該抗體或其抗原結合片段是44A8或其抗原結合片段、其嵌合抗體、或其人源化抗體，或它們的變體，該變體基本保留了其所源自的抗體或其抗原結合片段的生物學功能。In some preferred embodiments, the antibody or antigen-binding fragment thereof is 44A8 or its antigen-binding fragment, its chimeric antibody, or its humanized antibody, or a variant thereof, which substantially retains its The biological function of the derived antibody or its antigen-binding fragment.

在某些較佳的實施方案中，該抗體或其抗原結合片段具備以下生物學功能中的一項或多項： (a) 以0.1 μg/ml或更小的EC50結合人CLDN18.2； (b) 以0.1 μg/ml或更小的EC50結合小鼠CLDN18.2； (c) 不結合人CLDN18.1； (d) 藉由抗體依賴性細胞介導的細胞毒性（ADCC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (e) 藉由補體依賴的細胞毒性（CDC）來誘導殺傷表達人CLDN18.2的細胞（例如腫瘤細胞，如表達CLDN18.2的腫瘤細胞）； (f) 介導CLDN18.2內化至細胞（例如，腫瘤細胞）中，例如以如藉由FACS或流式細胞術所測量的至少10%（例如至少15%，至少20%或更多）的內化水準；該細胞在其表面表達CLDN18.2； (g) 在受試者中預防和/治療腫瘤（例如，表達CLDN18.2的腫瘤）。In some preferred embodiments, the antibody or antigen-binding fragment thereof has one or more of the following biological functions: (a) Binding to human CLDN 18.2 with an EC50 of 0.1 μg/ml or less; (b) Binding mouse CLDN18.2 with an EC50 of 0.1 μg/ml or less; (c) Does not combine with human CLDN 18.1; (d) Antibody-dependent cell-mediated cytotoxicity (ADCC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (e) Complement-dependent cytotoxicity (CDC) is used to induce the killing of cells expressing human CLDN 18.2 (e.g. tumor cells, such as tumor cells expressing CLDN 18.2); (f) Mediate the internalization of CLDN 18.2 into cells (e.g., tumor cells), for example at least 10% (e.g. at least 15%, at least 20% or more) as measured by FACS or flow cytometry The internalization level of the cell; the cell expresses CLDN18.2 on its surface; (g) Preventing and/treating tumors in subjects (for example, tumors expressing CLDN 18.2).

在某些較佳的實施方案中，該抗體或其抗原結合片段包含： (1) 如SEQ ID NO: 67所示的VH CDR1、如SEQ ID NO: 68所示的VH CDR2、如SEQ ID NO: 69所示的VH CDR3；如SEQ ID NO: 70所示的VL CDR1、如SEQ ID NO: 71所示的VL CDR2、如SEQ ID NO: 72所示的VL CDR3；或 (2) SEQ ID NO: 65所示的重鏈可變區中含有的3個CDR；以及，SEQ ID NO: 66所示的輕鏈可變區中含有的3個CDR。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: (1) VH CDR1 shown in SEQ ID NO: 67, VH CDR2 shown in SEQ ID NO: 68, VH CDR3 shown in SEQ ID NO: 69; VL CDR1 shown in SEQ ID NO: 70 , VL CDR2 as shown in SEQ ID NO: 71, VL CDR3 as shown in SEQ ID NO: 72; or (2) The 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 65; and the 3 CDRs contained in the light chain variable region shown in SEQ ID NO: 66.

在一個示例性實施方案中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 65所示的序列； (ii) 與SEQ ID NO: 65所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 65所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 66所示的序列； (v) 與SEQ ID NO: 66所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 66所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列。In an exemplary embodiment, the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 65; (ii) Compared with the sequence shown in SEQ ID NO: 65, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 65 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 66; (v) Compared with the sequence shown in SEQ ID NO: 66, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 66 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity.

在某些較佳的實施方案中，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 65所示的序列的VH和具有如SEQ ID NO: 66所示的序列的VL。In certain preferred embodiments, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 65 and VL having the sequence shown in SEQ ID NO: 66.

在某些較佳的實施方案中，本發明的抗體或其抗原結合片段可以進一步包含來源於哺乳動物（例如，鼠或人）免疫球蛋白的恆定區序列或其變體，該變體與其所源自的序列相比具有一或複數氨基酸的置換、缺失或添加。在某些較佳的實施方案中，該變體與其所源自的序列相比具有一或複數氨基酸的保守置換。In certain preferred embodiments, the antibody or antigen-binding fragment thereof of the present invention may further comprise a constant region sequence derived from a mammalian (for example, murine or human) immunoglobulin or a variant thereof, and the variant is Compared with the derived sequence, there are substitutions, deletions or additions of one or more amino acids. In certain preferred embodiments, the variant has conservative substitutions of one or more amino acids compared to the sequence from which it is derived.

在某些較佳的實施方案中，本發明的抗體或其抗原結合片段的重鏈包含人免疫球蛋白的重鏈恆定區（CH）或其變體，該變體與其所源自的序列相比具有一或複數氨基酸的置換、缺失或添加（例如，至多20個、至多15個、至多10個、或至多5個氨基酸的置換、缺失或添加；例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）；和/或，本發明的抗體或其抗原結合片段的輕鏈包含人免疫球蛋白的輕鏈恆定區（CL）或其變體，該變體與其所源自的序列相比具有至多20個氨基酸的保守置換（例如至多15個、至多10個、或至多5個氨基酸的保守置換；例如1個、2個、3個、4個或5個氨基酸的保守置換）。In certain preferred embodiments, the heavy chain of the antibody or antigen-binding fragment thereof of the present invention comprises the heavy chain constant region (CH) of human immunoglobulin or a variant thereof, which is similar to the sequence from which it is derived. Compared with substitutions, deletions, or additions with one or more amino acids (e.g., up to 20, up to 15, up to 10, or up to 5 amino acid substitutions, deletions, or additions; for example, 1, 2, 3, 4 Substitution, deletion or addition of one or five amino acids); and/or, The light chain of the antibody or antigen-binding fragment thereof of the present invention comprises the light chain constant region (CL) of human immunoglobulin or a variant thereof, which has conservative substitutions of up to 20 amino acids compared to the sequence from which it is derived ( For example, conservative substitutions of up to 15, up to 10, or up to 5 amino acids; for example, conservative substitutions of 1, 2, 3, 4, or 5 amino acids).

在某些較佳的實施方案中，該重鏈恆定區是IgG重鏈恆定區，例如IgG1、IgG2、IgG3或IgG4重鏈恆定區。在某些較佳的實施方案中，該重鏈恆定區是鼠IgG1、IgG2、IgG3或IgG4重鏈恆定區。在某些較佳的實施方案中，該重鏈恆定區是人IgG1、IgG2、IgG3或IgG4重鏈恆定區。在某些實施方案中，較佳地該重鏈恆定區是人IgG1或IgG4重鏈恆定區。In certain preferred embodiments, the heavy chain constant region is an IgG heavy chain constant region, such as an IgG1, IgG2, IgG3, or IgG4 heavy chain constant region. In certain preferred embodiments, the heavy chain constant region is a murine IgG1, IgG2, IgG3 or IgG4 heavy chain constant region. In certain preferred embodiments, the heavy chain constant region is a human IgG1, IgG2, IgG3 or IgG4 heavy chain constant region. In certain embodiments, preferably the heavy chain constant region is a human IgG1 or IgG4 heavy chain constant region.

在某些較佳的實施方案中，該輕鏈恆定區是κ輕鏈恆定區。在某些較佳的實施方案中，該輕鏈恆定區是鼠κ輕鏈恆定區。在某些較佳的實施方案中，該輕鏈恆定區是人κ輕鏈恆定區。In certain preferred embodiments, the light chain constant region is a kappa light chain constant region. In certain preferred embodiments, the light chain constant region is a murine kappa light chain constant region. In certain preferred embodiments, the light chain constant region is a human kappa light chain constant region.

在某些示例性實施方案中，本發明的抗體或其抗原結合片段包含SEQ ID NO: 81所示的重鏈恆定區（CH）；和/或，SEQ ID NO: 82所示的輕鏈恆定區（CL）。In certain exemplary embodiments, the antibody or antigen-binding fragment thereof of the present invention comprises the heavy chain constant region (CH) shown in SEQ ID NO: 81; and/or, the light chain constant region shown in SEQ ID NO: 82 District (CL).

在某些較佳的實施方案中，本發明的抗體是鼠源抗體、嵌合抗體、人源化抗體、雙特異性抗體或多特異性抗體。在某些較佳的實施方案中，本發明的抗原結合片段選自Fab、Fab’、(Fab’)₂ 、Fv、二硫鍵連接的Fv、scFv、雙抗體(diabody)和單域抗體(sdAb)。In certain preferred embodiments, the antibodies of the present invention are murine antibodies, chimeric antibodies, humanized antibodies, bispecific antibodies or multispecific antibodies. In certain preferred embodiments, the antigen-binding fragments of the present invention are selected from Fab, Fab', (Fab') ₂ , Fv, disulfide-linked Fv, scFv, diabody, and single domain antibody ( sdAb).

在本發明中，本發明的抗體或其抗原結合片段可以包括這樣的變體，該變體與其所源自的抗體或其抗原結合片段相比差異僅在於一或複數（例如，至多20個、至多15個、至多10個、或至多5個氨基酸的保守置換）氨基酸殘基的保守置換，或者與其所源自的抗體或其抗原結合片段具有至少85%、至少90%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性，且基本保留了其所源自的抗體或其抗原結合片段的上述生物學功能。抗體的製備In the present invention, the antibody or antigen-binding fragment thereof of the present invention may include a variant that differs from the antibody or antigen-binding fragment from which it is derived by only one or plural (for example, up to 20, Conservative substitution of at most 15, at most 10, or at most 5 amino acids) conservative substitution of amino acid residues, or at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity, and basically retains the above-mentioned biological functions of the antibody or antigen-binding fragment from which it is derived. Antibody preparation

本發明的抗體可以本領域已知的各種方法來製備，例如藉由基因工程重組技術來獲得。例如，藉由化學合成或PCR擴增獲得編碼本發明抗體的重鏈和輕鏈基因的DNA分子。將所得DNA分子***表達載體內，然後轉染宿主細胞。然後，在特定條件下培養轉染後的宿主細胞，並表達本發明的抗體。The antibody of the present invention can be prepared by various methods known in the art, for example, obtained by genetic engineering recombination technology. For example, DNA molecules encoding the heavy chain and light chain genes of the antibody of the present invention are obtained by chemical synthesis or PCR amplification. The resulting DNA molecule is inserted into the expression vector and then transfected into the host cell. Then, the transfected host cell is cultured under specific conditions, and the antibody of the present invention is expressed.

本發明的抗原結合片段可以藉由水解完整的抗體分子獲得（參見Morimoto et al., J. Biochem. Biophys. Methods 24:107-117 (1992) and Brennan et al., Science 229:81 (1985)）。另外，這些抗原結合片段也可以直接由重組宿主細胞產生（reviewed in Hudson, Curr. Opin. Immunol. 11: 548-557 (1999); Little et al., Immunol. Today, 21: 364-370 (2000)）。比如，Fab’片段可以直接從宿主細胞中獲得；可以將Fab’片段化學偶聯形成F(ab’)₂ 片段(Carter et al., Bio/Technology, 10: 163-167 (1992))。另外，Fv、Fab或F(ab’)₂ 片段也可以直接從重組宿主細胞培養液中直接分離得到。本領域的普通技術人員完全知曉製備這些抗原結合片段的其它技術。The antigen-binding fragments of the present invention can be obtained by hydrolyzing intact antibody molecules (see Morimoto et al., J. Biochem. Biophys. Methods 24:107-117 (1992) and Brennan et al., Science 229:81 (1985) ). In addition, these antigen-binding fragments can also be directly produced by recombinant host cells (reviewed in Hudson, Curr. Opin. Immunol. 11: 548-557 (1999); Little et al., Immunol. Today, 21: 364-370 (2000) )). For example, Fab' fragments can be obtained directly from host cells; Fab' fragments can be chemically coupled to form F(ab') ₂ fragments (Carter et al., Bio/Technology, 10: 163-167 (1992)). In addition, Fv, Fab or F(ab') ₂ fragments can also be directly isolated from the recombinant host cell culture medium. Those of ordinary skill in the art are fully aware of other techniques for preparing these antigen-binding fragments.

因此，在另一個方面，本發明提供了一種分離的核酸分子，其包含編碼本發明的抗體或其抗原結合片段，或其重鏈可變區和/或輕鏈可變區的核苷酸序列。在某些較佳的實施方案中，該分離的核酸分子編碼本發明的抗體或其抗原結合片段，或其重鏈可變區和/或輕鏈可變區。Therefore, in another aspect, the present invention provides an isolated nucleic acid molecule comprising a nucleotide sequence encoding the antibody or antigen-binding fragment thereof of the present invention, or its heavy chain variable region and/or light chain variable region . In certain preferred embodiments, the isolated nucleic acid molecule encodes the antibody or antigen-binding fragment thereof of the present invention, or the heavy chain variable region and/or light chain variable region thereof.

在另一個方面，本發明提供了一種載體（例如殖株載體或表達載體），其包含本發明的分離的核酸分子。在某些較佳的實施方案中，本發明的載體是例如質粒、黏粒、噬菌體等。在某些較佳的實施方案中，該載體能夠在受試者（例如哺乳動物，例如人）體內表達本發明的抗體或其抗原結合片段。In another aspect, the present invention provides a vector (such as a clone vector or an expression vector) comprising the isolated nucleic acid molecule of the present invention. In some preferred embodiments, the vectors of the present invention are, for example, plasmids, cosmids, bacteriophages and the like. In certain preferred embodiments, the vector is capable of expressing the antibody or antigen-binding fragment thereof of the present invention in a subject (such as a mammal, such as a human).

在另一個方面，本發明提供了一種宿主細胞，其包含本發明的分離的核酸分子或本發明的載體。此類宿主細胞包括但不限於，原核細胞例如大腸桿菌細胞，以及真核細胞例如酵母細胞、昆蟲細胞、植物細胞和動物細胞（如哺乳動物細胞，例如小鼠細胞、人細胞等）。在某些較佳的實施方案中，本發明的宿主細胞是哺乳動物細胞，例如CHO（例如CHO-K1、CHO-S、CHO DG44）。In another aspect, the invention provides a host cell comprising the isolated nucleic acid molecule of the invention or the vector of the invention. Such host cells include, but are not limited to, prokaryotic cells such as E. coli cells, and eukaryotic cells such as yeast cells, insect cells, plant cells, and animal cells (such as mammalian cells, such as mouse cells, human cells, etc.). In certain preferred embodiments, the host cell of the present invention is a mammalian cell, such as CHO (eg CHO-K1, CHO-S, CHO DG44).

在另一個方面，提供了製備本發明的抗體或其抗原結合片段的方法，其包括，在允許該抗體或其抗原結合片段表達的條件下，培養本發明的宿主細胞，和從培養的宿主細胞培養物中回收該抗體或其抗原結合片段。衍生的抗體In another aspect, there is provided a method for preparing the antibody or antigen-binding fragment thereof of the present invention, which comprises culturing the host cell of the present invention under conditions that allow the expression of the antibody or antigen-binding fragment thereof, and obtaining from the cultured host cell The antibody or its antigen-binding fragment is recovered from the culture. Derived antibodies

本發明的抗體或其抗原結合片段可進行衍生化，例如被連接至另一個分子（例如另一個多肽或蛋白）。通常，抗體或其抗原結合片段的衍生化（例如，標記）不會不利影響其對CLDN18.2（特別是人CLDN18.2）的結合。因此，本發明的抗體或其抗原結合片段還意欲包括此類衍生化的形式。例如，可以將本發明的抗體或其抗原結合片段功能性連接（藉由化學偶合、基因融合、非共價連接或其它方式）於一或複數其它分子基團，例如另一個抗體（例如，形成雙特異性抗體）、檢測試劑、藥用試劑，和/或能夠介導抗體或抗原結合片段與另一個分子結合的蛋白或多肽（例如，抗生物素蛋白或多組氨酸標籤）。此外，本發明的抗體或其抗原結合片段還可以用化學基團進行衍生，例如聚乙二醇（PEG）、甲基或乙基、或者糖基。這些基團可用於改善抗體的生物學特性，例如增加血清半衰期。The antibody or antigen-binding fragment thereof of the present invention may be derivatized, for example, linked to another molecule (for example, another polypeptide or protein). Generally, the derivatization (for example, labeling) of the antibody or its antigen-binding fragment will not adversely affect its binding to CLDN 18.2 (especially human CLDN 18.2). Therefore, the antibodies or antigen-binding fragments thereof of the present invention are also intended to include such derivatized forms. For example, the antibody or antigen-binding fragment thereof of the present invention can be functionally linked (by chemical coupling, gene fusion, non-covalent linkage or other means) to one or more other molecular groups, such as another antibody (for example, to form Bispecific antibodies), detection reagents, pharmaceutical reagents, and/or proteins or polypeptides capable of mediating the binding of an antibody or antigen-binding fragment to another molecule (for example, avidin or polyhistidine tag). In addition, the antibody or antigen-binding fragment thereof of the present invention can also be derivatized with chemical groups, such as polyethylene glycol (PEG), methyl or ethyl, or sugar groups. These groups can be used to improve the biological properties of antibodies, such as increasing serum half-life.

因此，在某些較佳的實施方案中，本發明的抗體或其抗原結合片段帶有標記。在某些較佳的實施方案中，本發明的抗體或其抗原結合片段帶有可檢測的標記，例如酶、放射性核素、螢光染料、發光物質（如化學發光物質）或生物素。本發明所述的可檢測的標記可以是可藉由螢光、光譜、光化學、生物化學、免疫學、電學、光學或化學手段檢測的任何物質。這類標記是本領域熟知的，其實例包括但不限於，酶（例如，辣根過氧化物酶、鹼性磷酸酶、β-半乳糖苷酶、脲酶、葡萄糖氧化酶，等）、放射性核素（例如，³ H、¹²⁵ I、³⁵ S、¹⁴ C或³² P）、螢光染料（例如，異硫氰酸螢光素（FITC）、螢光素、異硫氰酸四甲基羅丹明（TRITC）、藻紅蛋白（PE）、德克薩斯紅、羅丹明、量子點或花菁染料衍生物（例如Cy7、Alexa 750））、發光物質（例如化學發光物質，如吖啶酯類化合物）、磁珠（例如，Dynabeads^® ）、測熱標記物例如膠體金或有色玻璃或塑膠（例如，聚苯乙烯、聚丙烯、乳膠，等）珠、以及用於結合上述標記物修飾的親和素（例如，鏈黴親和素）的生物素。教導該標記物的使用的專利包括，但不限於，美國專利3,817,837；3,850,752；3,939,350；3,996,345；4,277,437；4,275,149；及4,366,241（全部藉由引用併入本文）。如上所述的可檢測的標記可藉由本領域已知的方法檢測。例如，放射性標記可使用攝影膠片或閃爍計算器檢測，螢光標記物可使用光檢測器檢測，以檢測發射的光。酶標記物一般藉由給酶提供底物及檢測藉由酶對底物的作用產生的反應產物來檢測，及測熱標記物藉由簡單視覺化著色標記物來檢測。在某些實施方案中，此類標記能夠適用於免疫學檢測（例如，酶聯免疫測定法、放射免疫測定法、螢光免疫測定法、化學發光免疫測定法等）。在某些實施方案中，可藉由不同長度的接頭（linker）將如上所述的可檢測的標記連接至本發明的抗體或其抗原結合片段，以降低潛在的位阻。雙特異性或多特異性分子Therefore, in certain preferred embodiments, the antibodies or antigen-binding fragments thereof of the present invention are labeled. In some preferred embodiments, the antibody or antigen-binding fragment thereof of the present invention bears a detectable label, such as an enzyme, a radionuclide, a fluorescent dye, a luminescent substance (such as a chemiluminescent substance), or biotin. The detectable label in the present invention can be any substance that can be detected by fluorescence, spectroscopy, photochemistry, biochemistry, immunology, electrical, optical or chemical means. Such labels are well-known in the art, and examples thereof include, but are not limited to, enzymes (for example, horseradish peroxidase, alkaline phosphatase, β-galactosidase, urease, glucose oxidase, etc.), radioactive nuclear (E.g., ³ H, ¹²⁵ I, ³⁵ S, ¹⁴ C, or ³² P), fluorescent dyes (e.g., fluorescein isothiocyanate (FITC), luciferin, tetramethylrhodamine isothiocyanate) (TRITC), phycoerythrin (PE), Texas red, rhodamine, quantum dots or cyanine dye derivatives (such as Cy7, Alexa 750)), luminescent substances (such as chemiluminescent substances, such as acridine esters) Compounds), magnetic beads (for example, Dynabeads ^® ), calorimetric markers such as colloidal gold or colored glass or plastic (for example, polystyrene, polypropylene, latex, etc.) beads, and affinity modified by combining the above-mentioned markers Biotin (for example, streptavidin). Patents teaching the use of this marker include, but are not limited to, US Patent Nos. 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241 (all incorporated herein by reference). The detectable label as described above can be detected by methods known in the art. For example, a radioactive label can be detected using photographic film or a scintillation calculator, and a fluorescent label can be detected using a light detector to detect the emitted light. Enzyme markers are generally detected by providing a substrate for the enzyme and detecting reaction products produced by the action of the enzyme on the substrate, and calorimetric markers are detected by simply visualizing colored markers. In certain embodiments, such labels can be suitable for immunological detection (eg, enzyme-linked immunoassay, radioimmunoassay, fluoroimmunoassay, chemiluminescence immunoassay, etc.). In some embodiments, the detectable label as described above can be connected to the antibody or antigen-binding fragment thereof of the present invention by linkers of different lengths to reduce potential steric hindrance. Bispecific or multispecific molecule

本發明的抗體或其抗原結合片段可用於形成雙特異性或多特異性分子。本發明的抗體或其抗原結合片段可以是雙特異性或多特異性分子的一部分，該雙特異性或多特異性分子包含具有不同於本發明的抗體或其抗原結合片段的結合特異性的第二功能模組（例如第二抗體），從而能夠結合至少兩個不同的結合位點和/或靶分子。例如，本發明的抗體或其抗原結合片段可連接至能夠特異性結合可用作聯合治療的潛在靶標的任何蛋白質的第二抗體或其抗原結合片段。為產生該雙特異性或多特異性分子，可以將本發明的抗體或其抗原結合片段連接（例如藉由化學偶聯、基因融合、非共價締合或其他方式）至一或複數其他結合分子（例如另外的抗體、抗體片段、肽或結合模擬物）。The antibodies or antigen-binding fragments thereof of the present invention can be used to form bispecific or multispecific molecules. The antibody or antigen-binding fragment thereof of the present invention may be a part of a bispecific or multispecific molecule, the bispecific or multispecific molecule comprising a second binding specificity that is different from that of the antibody or antigen-binding fragment thereof of the present invention. Two functional modules (such as a second antibody), which can bind at least two different binding sites and/or target molecules. For example, the antibody or antigen-binding fragment thereof of the present invention can be linked to a second antibody or antigen-binding fragment thereof that can specifically bind to any protein that can be used as a potential target for combination therapy. To produce the bispecific or multispecific molecule, the antibody or antigen-binding fragment of the present invention can be linked (for example, by chemical coupling, gene fusion, non-covalent association or other means) to one or more other bindings Molecules (eg, additional antibodies, antibody fragments, peptides, or binding mimetics).

因此，在另一個方面，本發明提供了一種雙特異性或多特異性分子，其包含本發明的抗體或其抗原結合片段。Therefore, in another aspect, the present invention provides a bispecific or multispecific molecule comprising the antibody or antigen-binding fragment thereof of the present invention.

在某些較佳的實施方案中，該雙特異性或多特異性分子特異性結合CLDN18.2，並且額外地特異性結合一或複數其他靶標。In certain preferred embodiments, the bispecific or multispecific molecule specifically binds to CLDN 18.2, and additionally specifically binds to one or more other targets.

在某些較佳的實施方案中，該雙特異性或多特異性分子還包含至少一種具有針對第二靶標的第二結合特異性的分子（例如第二抗體）。免疫綴合物In certain preferred embodiments, the bispecific or multispecific molecule further comprises at least one molecule having a second binding specificity for a second target (eg, a second antibody). Immunoconjugate

本發明的抗體或其抗原結合片段可以與治療劑連接以形成免疫綴合物。由於免疫綴合物具有選擇性遞送一種或多種治療劑至靶組織（例如與腫瘤相關的抗原，如表達CLDN18.2的腫瘤）的能力，因此，免疫綴合物可以提高本發明的抗體或其抗原結合片段在治療疾病（例如癌症）中的治療效力。The antibody or antigen-binding fragment thereof of the present invention can be linked to a therapeutic agent to form an immunoconjugate. Since immunoconjugates have the ability to selectively deliver one or more therapeutic agents to target tissues (for example, tumor-associated antigens, such as tumors expressing CLDN 18.2), immunoconjugates can improve the antibody of the present invention or its The therapeutic efficacy of antigen-binding fragments in the treatment of diseases such as cancer.

因此，在另一個方面，本發明提供了免疫綴合物，其包含本發明的抗體或其抗原結合片段以及連接於該抗體或其抗原結合片段的治療劑。Therefore, in another aspect, the present invention provides an immunoconjugate comprising the antibody or antigen-binding fragment thereof of the present invention and a therapeutic agent linked to the antibody or antigen-binding fragment thereof.

在某些較佳的實施方案中，該免疫綴合物是抗體-藥物偶聯物(ADC)。In certain preferred embodiments, the immunoconjugate is an antibody-drug conjugate (ADC).

在某些較佳的實施方案中，該治療劑是細胞毒劑。在本發明中，該細胞毒劑包括對細胞有害(例如殺傷細胞)的任何試劑。In certain preferred embodiments, the therapeutic agent is a cytotoxic agent. In the present invention, the cytotoxic agent includes any agent that is harmful to cells (e.g., kills cells).

在某些較佳的實施方案中，該治療劑選自烷化劑、有絲***抑制劑、抗腫瘤抗生素、抗代謝物、拓撲異構酶抑制劑、酪氨酸激酶抑制劑、放射性核素劑，及其任意組合。In certain preferred embodiments, the therapeutic agent is selected from alkylating agents, mitotic inhibitors, anti-tumor antibiotics, antimetabolites, topoisomerase inhibitors, tyrosine kinase inhibitors, radionuclide agents, And any combination.

可用於本發明的免疫綴合物的烷化劑的實例包括但不限於氮芥類（如雙氯乙基甲胺、苯丁酸氮芥、美法侖、環磷醯胺等）、乙烯亞胺類（如塞替呱等）、磺酸酯及多元醇類（如白消安、二溴甘露醇）、亞硝基脲類（如卡莫司汀、洛莫司汀等）、鉑類抗腫瘤劑（如順鉑、奧沙利鉑、卡鉑等）等。Examples of alkylating agents that can be used in the immunoconjugates of the present invention include, but are not limited to, nitrogen mustards (such as dichloroethyl methylamine, chlorambucil, melphalan, cyclophosphamide, etc.), ethylene Amines (such as cetigua, etc.), sulfonate and polyols (such as busulfan, dibromomannitol), nitrosoureas (such as carmustine, lomustine, etc.), platinum Anti-tumor agents (such as cisplatin, oxaliplatin, carboplatin, etc.), etc.

可用於本發明的免疫綴合物的有絲***抑制劑的實例包括但不限於美登素類（例如美登素、美登醇、美登醇的C-3酯等）、紫杉烷類（例如多西他賽、紫杉醇或奈米顆粒紫杉醇等）、長春花生物鹼類（例如硫酸長春地辛、長春新鹼、長春花鹼或長春瑞濱等）Examples of mitosis inhibitors that can be used in the immunoconjugates of the present invention include, but are not limited to, maytansinoids (such as maytansine, maytansinol, C-3 esters of maytansinol, etc.), taxanes (such as Docetaxel, paclitaxel or nanoparticle paclitaxel, etc.), vinca alkaloids (such as vindesine sulfate, vincristine, vinblastine or vinorelbine, etc.)

可用於本發明的免疫綴合物的抗腫瘤抗生素的實例包括但不限於放線菌素、蒽環類抗生素（例如柔紅黴素、阿黴素、表柔比星、伊達比星等）、卡利奇黴素、倍癌黴素等。Examples of anti-tumor antibiotics that can be used in the immunoconjugates of the present invention include, but are not limited to, actinomycin, anthracycline antibiotics (such as daunorubicin, doxorubicin, epirubicin, idarubicin, etc.), Lichomycin, becinomycin and so on.

可用於本發明的免疫綴合物的抗代謝物的實例包括但不限於葉酸拮抗劑（例如甲氨蝶呤等）、嘧啶拮抗劑（例如5-氟尿嘧啶、氟尿苷、阿糖胞苷、卡培他濱、吉西他濱等）、嘌呤拮抗劑（例如6-巰基嘌呤、6-硫鳥嘌呤等）、腺苷脫氨酶抑制劑（例如克拉屈濱、氟達拉濱、奈拉濱、噴司他丁等）。Examples of antimetabolites that can be used in the immunoconjugates of the present invention include, but are not limited to, folate antagonists (e.g. methotrexate, etc.), pyrimidine antagonists (e.g. 5-fluorouracil, fluorouridine, cytarabine, card Peitabine, gemcitabine, etc.), purine antagonists (such as 6-mercaptopurine, 6-thioguanine, etc.), adenosine deaminase inhibitors (such as cladribine, fludarabine, nelarabine, pens Tatin etc.).

可用於本發明的免疫綴合物的拓撲異構酶抑制劑的實例包括但不限於（喜樹鹼類及其衍生物（例如伊立替康、托泊替康等）、安吖啶、道諾黴素、阿黴素、表鬼臼毒素類、玫瑰樹鹼類、表柔比星、依託泊苷、丙亞胺、替尼泊苷等。Examples of topoisomerase inhibitors that can be used in the immunoconjugates of the present invention include but are not limited to (camptothecins and their derivatives (for example, irinotecan, topotecan, etc.), amsacrine, daunor Amycin, doxorubicin, epipodophyllotoxin, ellipticine, epirubicin, etoposide, propylimine, teniposide, etc.

可用於本發明的免疫綴合物的酪氨酸激酶抑制劑的實例包括但不限於阿西替尼、博舒替尼、西地尼布、達沙替尼、厄洛替尼、吉非替尼、伊馬替尼、拉帕替尼、來妥替尼、尼洛替尼、司馬沙尼、舒尼替尼、凡德他尼等。Examples of tyrosine kinase inhibitors that can be used in the immunoconjugates of the present invention include, but are not limited to, axitinib, bosutinib, cediranib, dasatinib, erlotinib, gefitin Ni, imatinib, lapatinib, letutinib, nilotinib, simazanib, sunitinib, vandetanib, etc.

可用於本發明的免疫綴合物的放射性核素劑的實例包括但不限於I¹³¹ 、In¹¹¹ 、Y⁹⁰ 、Lu¹⁷⁷ 等。Examples of radionuclide agents that can be used in the immunoconjugates of the present invention include, but are not limited to, I ¹³¹ , In ¹¹¹ , Y ⁹⁰ , Lu ¹⁷⁷ and the like.

在某些實例性實施方案中，該治療劑選自鉑類抗腫瘤劑、蒽環類抗生素、紫杉烷類化合物、核苷類似物、喜樹鹼類化合物，及其類似物或同系物，及其任意組合。In certain exemplary embodiments, the therapeutic agent is selected from platinum anti-tumor agents, anthracycline antibiotics, taxanes, nucleoside analogs, camptothecins, and analogs or homologs thereof, And any combination.

在某些較佳的實施方案中，本發明的抗體或其抗原結合片段任選地藉由接頭與該治療劑綴合。In certain preferred embodiments, the antibody or antigen-binding fragment thereof of the present invention is optionally conjugated to the therapeutic agent via a linker.

在本發明中，使用本領域現有的接頭技術可以將細胞毒劑偶聯至本發明的抗體或其抗原結合片段。已經用於將細胞毒劑偶聯至抗體的接頭類型的實例包括但不限於腙、硫醚、酯、二硫化物和含肽的接頭。可以選擇例如易於在溶酶體隔室內被低pH切割或易於被蛋白酶（例如優先在腫瘤組織中表達的蛋白酶，如組織蛋白酶，諸如組織蛋白酶B、C、D）切割的接頭。In the present invention, the cytotoxic agent can be coupled to the antibody or antigen-binding fragment thereof of the present invention by using linker technology available in the art. Examples of the types of linkers that have been used to couple cytotoxic agents to antibodies include, but are not limited to, hydrazones, thioethers, esters, disulfides, and peptide-containing linkers. For example, linkers that are easy to be cleaved by low pH in the lysosomal compartment or proteases (eg, proteases that are preferentially expressed in tumor tissues, such as cathepsins, such as cathepsins B, C, D) can be selected.

關於細胞毒劑的類型、接頭及將治療劑偶聯至抗體的方法的進一步討論還可參見Saito，G.等人(2003)Adv.Drug Deliv.Rev.55：199-215；Trail，P.A.等人(2003)Cancer Immunol.Immunother.52：328-337；Payne，G.(2003)Cancer Cell 3：207-212；Allen，T.M.(2002)Nat.Rev.Cancer 2：750-763；Pastan，I.and Kreitman，R.J.(2002)Curr.Opin.Investig.Drugs 3：1089-1091；Senter，P.D.and Springer，C.J.(2001)Adv.Drug Deliv.Rev.53：247-264。嵌合抗原受體For further discussion on the types of cytotoxic agents, linkers, and methods for coupling therapeutic agents to antibodies, see Saito, G. et al. (2003) Adv. Drug Deliv. Rev. 55: 199-215; Trail, PA, et al. (2003) Cancer Immunol. Immunother. 52: 328-337; Payne, G. (2003) Cancer Cell 3: 207-212; Allen, TM (2002) Nat. Rev. Cancer 2: 750-763; Pastan, I. and Kreitman, RJ (2002) Curr. Opin. Investig. Drugs 3: 1089-1091; Senter, PD and Springer, CJ (2001) Adv. Drug Deliv. Rev. 53: 247-264. Chimeric antigen receptor

本發明的抗體或其抗原結合片段可用於構建嵌合抗原受體(CAR)，該CAR包含特異性結合CLDN18.2的、與跨膜結構域連接的、與一或複數細胞內T細胞訊號結構域連接的細胞外抗原結合結構域（例如，scFv）。該細胞內T細胞訊號結構域可包括，例如T細胞受體訊號結構域、T細胞共刺激訊號結構域或其組合。T細胞受體訊號結構域是指CAR中包含T細胞受體的細胞內結構域(例如CD3ζ蛋白的細胞內部分)的部分。共刺激訊號結構域是指CAR中包含共刺激分子的細胞內結構域的部分，該共刺激分子是除了淋巴細胞對抗原的高效應答所需的抗原受體或其配體以外的細胞表面分子。The antibody or antigen-binding fragment thereof of the present invention can be used to construct a chimeric antigen receptor (CAR), which comprises a T cell signaling structure that specifically binds to CLDN 18.2, is connected to a transmembrane domain, and is connected to one or more intracellular T cells Domain-linked extracellular antigen binding domain (eg, scFv). The intracellular T cell signaling domain may include, for example, a T cell receptor signaling domain, a T cell costimulatory signaling domain, or a combination thereof. The T cell receptor signaling domain refers to the part of the CAR that contains the intracellular domain of the T cell receptor (for example, the intracellular part of the CD3ζ protein). The costimulatory signal domain refers to the part of the intracellular domain of a costimulatory molecule in the CAR. The costimulatory molecule is a cell surface molecule other than the antigen receptor or its ligand required for the efficient response of lymphocytes to the antigen.

本發明的CAR的特徵包括其以非MHC限制的方式將T-細胞特異性和反應性指向表達CLDN18.2的細胞（例如腫瘤細胞）的能力。非MHC限制的CLDN18.2識別能力賦予表達本發明的CAR的T細胞獨立於抗原加工而識別抗原的能力。The features of the CAR of the present invention include its ability to direct T-cell specificity and reactivity to cells expressing CLDN 18.2 (such as tumor cells) in a non-MHC-restricted manner. The non-MHC-restricted CLDN18.2 recognition ability confers the ability of T cells expressing the CAR of the present invention to recognize antigen independently of antigen processing.

因此，在另一個方面，本發明提供了一種嵌合抗原受體(CAR)，其包含本發明的抗體或其抗原結合片段的抗原結合結構域。Therefore, in another aspect, the present invention provides a chimeric antigen receptor (CAR) comprising the antigen-binding domain of the antibody or antigen-binding fragment thereof of the present invention.

在某些較佳的實施方案中，該抗原結合結構域包含本發明的抗體或其抗原結合片段的重鏈可變區和輕鏈可變區。In certain preferred embodiments, the antigen-binding domain comprises the heavy chain variable region and the light chain variable region of the antibody or antigen-binding fragment thereof of the present invention.

在某些較佳的實施方案中，該抗原結合結構域是scFv。In certain preferred embodiments, the antigen binding domain is a scFv.

在某些較佳的實施方案中，該嵌合抗原受體包含本發明抗體的抗原結合片段（例如scFv）。In certain preferred embodiments, the chimeric antigen receptor comprises an antigen-binding fragment (eg, scFv) of the antibody of the invention.

在某些較佳的實施方案中，該嵌合抗原受體由免疫效應細胞（例如T細胞）所表達。In certain preferred embodiments, the chimeric antigen receptor is expressed by immune effector cells (such as T cells).

在某些較佳的實施方案中，在該CAR的抗原結合結構域與跨膜結構域之間，可存在包含多肽序列的間隔結構域。該間隔結構域可包含多達300個氨基酸，較佳10至100個氨基酸，並且最較佳25至50個氨基酸。在一些實施方案中，該間隔結構域可包含免疫球蛋白結構域，例如人免疫球蛋白序列。在某些示例性實施方案中，該免疫球蛋白結構域包含免疫球蛋白CH2和CH3結構域序列。在此類實施方案中，不受特定理論的約束，認為CH2和CH3結構域使該CAR的抗原結合結構域從表達CAR的細胞的膜延伸出去，並且可更精確地模擬天然TCR的大小和結構域結構。In certain preferred embodiments, between the antigen binding domain and the transmembrane domain of the CAR, there may be a spacer domain containing a polypeptide sequence. The spacer domain may contain up to 300 amino acids, preferably 10 to 100 amino acids, and most preferably 25 to 50 amino acids. In some embodiments, the spacer domain may comprise an immunoglobulin domain, such as a human immunoglobulin sequence. In certain exemplary embodiments, the immunoglobulin domain comprises immunoglobulin CH2 and CH3 domain sequences. In such embodiments, without being bound by a specific theory, it is believed that the CH2 and CH3 domains allow the antigen-binding domain of the CAR to extend from the membrane of the CAR-expressing cell, and can more accurately mimic the size and structure of the natural TCR Domain structure.

在某些較佳的實施方案中，該跨膜結構域可衍生自天然來源或合成來源。在此類實施方案中，該結構域可衍生自任何膜結合的或跨膜的蛋白質。可用於本發明的CAR中的示例性跨膜結構域可包含至少T細胞受體的α、β或ζ鏈的跨膜區，該T細胞受體可以選自CD28、CD3ε、CD45、CD4、CD5、CDS、CD9、CD16、CD22、CD33、CD37、CD64、CD80、CD86、CD134、CD137、CD154。或者，該跨膜結構域可為合成的，在這種情況下，其將主要包含疏水殘基例如亮氨酸和纈氨酸。In certain preferred embodiments, the transmembrane domain can be derived from natural sources or synthetic sources. In such embodiments, the domain can be derived from any membrane-bound or transmembrane protein. Exemplary transmembrane domains that can be used in the CAR of the present invention may comprise at least the transmembrane region of the α, β, or ζ chain of a T cell receptor, which may be selected from CD28, CD3ε, CD45, CD4, CD5 , CDS, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, CD154. Alternatively, the transmembrane domain may be synthetic, in which case it will mainly contain hydrophobic residues such as leucine and valine.

在某些示例性實施方案中，該跨膜結構域包含T細胞受體的跨膜結構域，例如CD8跨膜結構域。In certain exemplary embodiments, the transmembrane domain comprises a transmembrane domain of a T cell receptor, such as a CD8 transmembrane domain.

在某些示例性實施方案中，該跨膜結構域包含T細胞共刺激分子（例如CD137或CD28）的跨膜結構域。In certain exemplary embodiments, the transmembrane domain comprises the transmembrane domain of a T cell costimulatory molecule (eg, CD137 or CD28).

在某些較佳的實施方案中，可用於在該CAR中使用的細胞內T細胞結構域的實例包括T細胞受體(TCR)的胞質序列和共刺激分子，該T細胞受體(TCR)的胞質序列和共刺激分子在抗原受體接合後協力啟動訊號轉導，以及這些序列和任何具有相同功能性能力的合成序列的任何衍生物或變體。In certain preferred embodiments, examples of intracellular T cell domains that can be used in the CAR include the cytoplasmic sequence of the T cell receptor (TCR) and costimulatory molecules, the T cell receptor (TCR) The cytoplasmic sequence of) and the costimulatory molecule cooperate to initiate signal transduction after the antigen receptor is engaged, and any derivative or variant of these sequences and any synthetic sequence with the same functional ability.

在某些較佳的實施方案中，該CAR的細胞內區可包含以刺激的方式起作用的初級胞質訊號序列，其可包含被稱為基於免疫受體酪氨酸的活化基序或ITAM的訊號基序。可包含於該CAR中的包含初級胞質訊號序列的ITAM的實例包括來自CD3ζ、FcRγ、FcRβ、CD3γ、CD3δ、CD3ε、CDS、CD22、CD79a、CD79b、和CD66d蛋白的那些。In some preferred embodiments, the intracellular region of the CAR may include a primary cytoplasmic signal sequence that acts in a stimulating manner, and it may include an activation motif called an immunoreceptor tyrosine-based activation motif or ITAM的signal motifs. Examples of ITAMs containing primary cytoplasmic signal sequences that can be included in the CAR include those from CD3ζ, FcRγ, FcRβ, CD3γ, CD3δ, CD3ε, CDS, CD22, CD79a, CD79b, and CD66d proteins.

在某些較佳的實施方案中，該CAR的細胞內區可包含含有初級胞質訊號結構域(例如CD3ζ)本身或與可用於CAR的環境中的任何其他期望的胞質結構域組合的ITAM。例如，該CAR的胞質結構域包含CD3ζ鏈部分和細胞內共刺激訊號結構域。該共刺激訊號結構域是指該CAR的包含共刺激分子的細胞內結構域的部分。共刺激分子是除了淋巴細胞對抗原的高效應答所需的抗原受體或其配體以外的細胞表面分子。這樣的分子的實例包括CD27、CD28、4-1BB(CD137)、OX40(CD134)、CD30、CD40、PD-1、ICOS、淋巴細胞功能相關抗原1(LFA-1)、CD2、CD7、LIGHT、NKG2C和B7-H3。In certain preferred embodiments, the intracellular region of the CAR may comprise an ITAM containing a primary cytoplasmic signaling domain (such as CD3ζ) itself or in combination with any other desired cytoplasmic domain that can be used in the CAR environment . For example, the cytoplasmic domain of the CAR contains the CD3ζ chain part and the intracellular co-stimulatory signal domain. The costimulatory signal domain refers to the part of the intracellular domain of the CAR that contains costimulatory molecules. Co-stimulatory molecules are cell surface molecules other than antigen receptors or their ligands required for efficient response of lymphocytes to antigens. Examples of such molecules include CD27, CD28, 4-1BB (CD137), OX40 (CD134), CD30, CD40, PD-1, ICOS, lymphocyte function associated antigen 1 (LFA-1), CD2, CD7, LIGHT, NKG2C and B7-H3.

在某些較佳的實施方案中，該CAR可包含CD3ζ訊號結構域、CD8訊號結構域、CD28訊號結構域、CD137訊號結構域或其任意組合。該一或複數T細胞訊號結構域在CAR上的順序可由本領域技術人員根據需要而改變。In some preferred embodiments, the CAR may comprise a CD3ζ signal domain, a CD8 signal domain, a CD28 signal domain, a CD137 signal domain, or any combination thereof. The order of the one or more T cell signal domains on the CAR can be changed by those skilled in the art as needed.

產生嵌合抗原受體、包含這樣的受體的T細胞的方法以及它們的用途(例如，用於治療癌症)是本領域已知的，其詳細描述可參見，例如，Brentjens等人, 2010, Molecular Therapy, 18:4,666-668；Morgan等人, 2010, Molecular Therapy, published online February 23, 2010, 第1-9頁；Till等人, 2008, Blood, 112:2261-2271；Park等人, Trends Biotechnol.,29:550-557,2011；Grupp等人, NEnglJMed., 368:1509-1518,2013；Han等人, J.Hematol Oncol.,6:47,2013；PCT專利公開文本WO2012/079000、WO2013/126726；和U.S.專利公開文本2012/0213783，其全部藉由引用整體併入本文)。例如，可將編碼本發明的嵌合抗原結合受體的核酸分子包含於用於在宿主細胞例如T細胞中表達的表達載體(例如，慢病毒載體)中，以製造該CAR。在某些實例性實施方案中，使用該嵌合抗原受體的方法包括從受試者分離T細胞，採用編碼嵌合抗原受體的表達載體(例如慢病毒載體)轉化T細胞，以及將表達該嵌合抗原受體的工程化的T細胞施用至該受試者以進行治療，例如用於治療該受試者中的腫瘤。Methods for generating chimeric antigen receptors, T cells containing such receptors, and their uses (for example, for the treatment of cancer) are known in the art, and a detailed description can be found in, for example, Brentjens et al., 2010, Molecular Therapy, 18:4,666-668; Morgan et al., 2010, Molecular Therapy, published online February 23, 2010, pages 1-9; Till et al., 2008, Blood, 112:2261-2271; Park et al., Trends Biotechnol., 29:550-557, 2011; Grupp et al., NEnglJMed., 368:1509-1518, 2013; Han et al., J. Hematol Oncol., 6:47, 2013; PCT Patent Publication WO2012/079000, WO2013/126726; and US Patent Publication 2012/0213783, all of which are incorporated herein by reference in their entirety). For example, the nucleic acid molecule encoding the chimeric antigen-binding receptor of the present invention can be included in an expression vector (e.g., lentiviral vector) for expression in a host cell, such as a T cell, to manufacture the CAR. In certain exemplary embodiments, the method of using the chimeric antigen receptor includes isolating T cells from a subject, transforming the T cells with an expression vector (such as a lentiviral vector) encoding the chimeric antigen receptor, and expressing The engineered T cells of the chimeric antigen receptor are administered to the subject for treatment, for example, to treat a tumor in the subject.

因此，在另一個方面，本發明提供了一種分離的核酸分子，其包含編碼本發明的嵌合抗原受體的核苷酸序列。在某些較佳的實施方案中，該分離的核酸分子編碼本發明的嵌合抗原受體。Therefore, in another aspect, the present invention provides an isolated nucleic acid molecule comprising a nucleotide sequence encoding the chimeric antigen receptor of the present invention. In certain preferred embodiments, the isolated nucleic acid molecule encodes the chimeric antigen receptor of the invention.

在另一個方面，本發明提供了一種載體（例如殖株載體或表達載體），其包含如上所述的分離的核酸分子。在某些較佳的實施方案中，本發明的載體例如是質粒。In another aspect, the present invention provides a vector (such as a clone vector or an expression vector) comprising the isolated nucleic acid molecule as described above. In certain preferred embodiments, the vector of the present invention is, for example, a plasmid.

在另一個方面，本發明提供了一種宿主細胞，其包含如上所述的分離的核酸分子或載體。在某些較佳的實施方案中，所述宿主細胞是T細胞。在某些較佳的實施方案中，所述宿主細胞是嵌合抗原受體T細胞（CAR-T）。治療方法和藥物組合物In another aspect, the present invention provides a host cell comprising the isolated nucleic acid molecule or vector as described above. In certain preferred embodiments, the host cell is a T cell. In certain preferred embodiments, the host cell is a chimeric antigen receptor T cell (CAR-T). Treatment method and pharmaceutical composition

本發明的抗體或其抗原結合片段可藉由結合CLDN18.2誘導ADCC和/或CDC從而殺傷細胞，從而能夠用於預防和/或治療腫瘤。The antibody or antigen-binding fragment thereof of the present invention can induce ADCC and/or CDC by binding to CLDN 18.2 to kill cells, and thus can be used to prevent and/or treat tumors.

因此，在另一個方面，本發明提供了一種藥物組合物，其含有本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、或免疫綴合物，以及藥學上可接受的載體和/或賦形劑。Therefore, in another aspect, the present invention provides a pharmaceutical composition containing the antibody or antigen-binding fragment thereof, bispecific or multispecific molecule, or immunoconjugate of the present invention, and a pharmaceutically acceptable carrier And/or excipients.

在某些較佳的實施方案中，該藥物組合物還可以包含另外的藥學活性劑。In certain preferred embodiments, the pharmaceutical composition may also contain additional pharmaceutically active agents.

在某些較佳的實施方案中，所述另外的藥學活性劑是具有抗腫瘤活性的藥物，例如烷化劑、有絲***抑制劑、抗腫瘤抗生素、抗代謝物、拓撲異構酶抑制劑、酪氨酸激酶抑制劑、放射性核素劑、放射增敏劑（例如吉西他濱、5-氟尿嘧啶、紫杉烷、順鉑等）、抗血管產生劑、細胞因數（例如GM-CSF、IL-7、IL-12、IL-15、IL-18、IL-21等）、分子靶向藥物（例如，CD20抗體如利妥昔單抗、Her2抗體如曲妥珠單抗、VEGF抗體如貝伐珠單抗、EGFR抗體如西妥昔單抗等）、免疫檢查點抑制劑（例如，PD-1抗體、PD-L1抗體、CTLA-4抗體、LAG-3抗體等）、溶瘤病毒等。In some preferred embodiments, the additional pharmacologically active agent is a drug with anti-tumor activity, such as alkylating agents, mitotic inhibitors, anti-tumor antibiotics, antimetabolites, topoisomerase inhibitors, tyrosine Acid kinase inhibitors, radionuclide agents, radiosensitizers (such as gemcitabine, 5-fluorouracil, taxane, cisplatin, etc.), antiangiogenic agents, cytokines (such as GM-CSF, IL-7, IL -12, IL-15, IL-18, IL-21, etc.), molecular targeted drugs (for example, CD20 antibody such as rituximab, Her2 antibody such as trastuzumab, VEGF antibody such as bevacizumab , EGFR antibodies such as cetuximab, etc.), immune checkpoint inhibitors (for example, PD-1 antibody, PD-L1 antibody, CTLA-4 antibody, LAG-3 antibody, etc.), oncolytic virus, etc.

在某些較佳的實施方案中，在該藥物組合物中，本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、或免疫綴合物與所述另外的藥學活性劑作為分離的組分或作為同一組合物的組分提供。因此，本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、或免疫綴合物與所述另外的藥學活性劑可以同時、分開或相繼施用。In certain preferred embodiments, in the pharmaceutical composition, the antibody or antigen-binding fragment, bispecific or multispecific molecule, or immunoconjugate of the present invention and the other pharmaceutically active agent are used as Separate components or provided as components of the same composition. Therefore, the antibody or antigen-binding fragment, bispecific or multispecific molecule, or immunoconjugate of the present invention and the additional pharmaceutically active agent can be administered simultaneously, separately or sequentially.

在某些示例性實施方案中，該藥物組合物包含無菌可注射液體（如水性或非水性懸浮液或溶液）。在某些示例性實施方案中，此類無菌可注射液體選自注射用水(WFI)、抑菌性注射用水(BWFI)、氯化鈉溶液（例如0.9% (w/v)NaCl）、葡萄糖溶液（例如5%葡萄糖）、含有表面活性劑的溶液（例如0.01%聚山梨醇20）、pH緩衝溶液（例如磷酸鹽緩衝溶液）、Ringer氏溶液及其任意組合。In certain exemplary embodiments, the pharmaceutical composition comprises a sterile injectable liquid (such as an aqueous or non-aqueous suspension or solution). In certain exemplary embodiments, such sterile injectable liquid is selected from the group consisting of water for injection (WFI), bacteriostatic water for injection (BWFI), sodium chloride solution (eg 0.9% (w/v) NaCl), glucose solution (Such as 5% glucose), a solution containing a surfactant (such as 0.01% polysorbate 20), a pH buffer solution (such as a phosphate buffer solution), Ringer's solution, and any combination thereof.

在另一個方面，本發明提供了一種降低CLDN18.2在細胞表面的表達水準的方法，其包括將該細胞與本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、或藥物組合物接觸，使得CLDN18.2在該細胞表面的表達水準降低；其中，該細胞在其表面表達CLDN18.2。In another aspect, the present invention provides a method for reducing the expression level of CLDN18.2 on the cell surface, which comprises combining the cell with the antibody or antigen-binding fragment thereof, bispecific or multispecific molecule, and immunoconjugate of the present invention. The contact with the compound or the pharmaceutical composition reduces the expression level of CLDN18.2 on the surface of the cell; wherein, the cell expresses CLDN18.2 on its surface.

在某些較佳的實施方案中，該細胞是表達CLDN18.2的腫瘤細胞。In certain preferred embodiments, the cell is a tumor cell expressing CLDN 18.2.

在某些較佳的實施方案中，該方法用於在體外降低CLDN18.2在細胞表面的表達水準用於非診斷目的。In some preferred embodiments, the method is used to reduce the expression level of CLDN 18.2 on the cell surface in vitro for non-diagnostic purposes.

在另一個方面，提供了本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、或藥物組合物在製備藥物中的用途，該藥物用於降低CLDN18.2在細胞表面的表達水準。In another aspect, there is provided the use of the antibody or antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, or pharmaceutical composition of the present invention in the preparation of a medicament for reducing CLDN 18.2 The level of expression on the cell surface.

在另一個方面，提供了本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、或藥物組合物，其用於降低CLDN18.2在細胞表面的表達水準。In another aspect, the antibody or antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, or pharmaceutical composition of the present invention is provided, which is used to reduce the expression level of CLDN 18.2 on the cell surface.

在另一個方面，本發明提供了一種抑制表達CLDN18.2的腫瘤細胞生長和/或殺傷該腫瘤細胞的方法，其包括將該腫瘤細胞與有效量的本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如嵌合抗原受體T細胞（CAR-T））接觸。In another aspect, the present invention provides a method for inhibiting the growth of tumor cells expressing CLDN 18.2 and/or killing the tumor cells, which comprises combining the tumor cells with an effective amount of the antibody of the present invention or its antigen-binding fragment, double Contact with specific or multispecific molecules, immunoconjugates, pharmaceutical compositions, chimeric antigen receptors, or host cells expressing the chimeric antigen receptors (such as chimeric antigen receptor T cells (CAR-T)).

該方法可以用於治療目的，或者非治療目的。在某些較佳的實施方案中，該方法可以用於非治療目的，該方法用於在體外抑制表達CLDN18.2的腫瘤細胞生長和/或殺傷該腫瘤細胞。This method can be used for therapeutic or non-therapeutic purposes. In certain preferred embodiments, the method can be used for non-therapeutic purposes. The method is used to inhibit the growth of tumor cells expressing CLDN 18.2 and/or kill the tumor cells in vitro.

在另一個方面，提供了本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如嵌合抗原受體T細胞（CAR-T））在製備藥物中的用途，該藥物用於抑制表達CLDN18.2的腫瘤細胞生長和/或殺傷該腫瘤細胞。In another aspect, the antibody or antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or host expressing the chimeric antigen receptor of the present invention is provided Use of cells (such as chimeric antigen receptor T cells (CAR-T)) in the preparation of medicines for inhibiting the growth of tumor cells expressing CLDN 18.2 and/or killing the tumor cells.

在另一個方面，提供了本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如嵌合抗原受體T細胞（CAR-T）），其用於抑制表達CLDN18.2的腫瘤細胞生長和/或殺傷該腫瘤細胞。In another aspect, the antibody or antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or host expressing the chimeric antigen receptor of the present invention is provided Cells (such as chimeric antigen receptor T cells (CAR-T)), which are used to inhibit the growth of tumor cells expressing CLDN 18.2 and/or kill the tumor cells.

在另一個方面，本發明提供了一種用於在受試者（例如人）中預防和/或治療腫瘤的方法，該方法包括向有此需要的受試者施用有效量的本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如嵌合抗原受體T細胞（CAR-T））。In another aspect, the present invention provides a method for preventing and/or treating tumors in a subject (such as a human), the method comprising administering to a subject in need thereof an effective amount of the antibody of the present invention or Its antigen-binding fragments, bispecific or multispecific molecules, immunoconjugates, pharmaceutical compositions, chimeric antigen receptors or host cells expressing the chimeric antigen receptors (such as chimeric antigen receptor T cells (CAR -T)).

在某些較佳的實施方案中，該腫瘤涉及表達CLDN18.2的腫瘤細胞。在某些較佳的實施方案中，該CLDN18.2在該腫瘤細胞表面上表達。In certain preferred embodiments, the tumor involves tumor cells expressing CLDN 18.2. In certain preferred embodiments, the CLDN 18.2 is expressed on the surface of the tumor cell.

在某些較佳的實施方案中，該腫瘤表達CLDN18.2。In certain preferred embodiments, the tumor expresses CLDN 18.2.

在某些較佳的實施方案中，該腫瘤選自胃癌、食道癌、胰腺癌、支氣管癌、非小細胞肺癌、乳腺癌、耳鼻喉(ENT)癌、卵巢癌、結腸癌、肝癌、頭頸癌、膽囊癌及其轉移癌（例如，胃癌轉移，如Krukenberg瘤、腹膜轉移或淋巴結轉移）。In certain preferred embodiments, the tumor is selected from gastric cancer, esophageal cancer, pancreatic cancer, bronchial cancer, non-small cell lung cancer, breast cancer, ear, nose and throat (ENT) cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer , Gallbladder cancer and its metastases (for example, gastric cancer metastasis, such as Krukenberg tumor, peritoneal metastasis or lymph node metastasis).

在某些較佳的實施方案中，將本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如嵌合抗原受體T細胞（CAR-T））與另外的具有抗腫瘤活性的藥物聯合使用。這種另外的具有抗腫瘤活性的藥物可以在施用該抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如CAR-T）之前、同時或之後施用。In some preferred embodiments, the antibody or antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or expression of the chimeric antigen of the present invention The recipient's host cells (such as chimeric antigen receptor T cells (CAR-T)) are used in combination with another drug with anti-tumor activity. This other drug with anti-tumor activity can be used to administer the antibody or its antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor, or express the chimeric antigen. The recipient's host cell (eg CAR-T) is administered before, at the same time or afterwards.

在某些較佳的實施方案中，將本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如CAR-T）與額外的療法組合施用。這種額外的療法可以是已知用於腫瘤的任何療法，例如手術、化學治療、放射治療、靶向治療、免疫治療、激素治療、基因治療或姑息治療。這種額外的療法可以在施用本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如CAR-T）之前、同時或之後施用。In some preferred embodiments, the antibody or antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or expression of the chimeric antigen of the present invention The recipient's host cells (eg CAR-T) are administered in combination with additional therapies. This additional therapy can be any therapy known for tumors, such as surgery, chemotherapy, radiation therapy, targeted therapy, immunotherapy, hormone therapy, gene therapy, or palliative therapy. This additional therapy can be used in the administration of the antibody or antigen-binding fragment, bispecific or multispecific molecule of the present invention, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or expression of the chimeric antigen receptor. The host cell (e.g. CAR-T) is administered before, simultaneously or afterwards.

本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如T細胞）可以配製成醫學領域已知的任何劑型，例如，片劑、丸劑、混懸劑、乳劑、溶液、凝膠劑、膠囊劑、粉劑、顆粒劑、酏劑、錠劑、栓劑、注射劑（包括注射液、注射用無菌粉末與注射用濃溶液）、吸入劑、噴霧劑等。較佳劑型取決於預期的給藥方式和治療用途。本發明的藥物組合物應當是無菌的並在生產和儲存條件下穩定。一種較佳的劑型是注射劑。此類注射劑可以是無菌注射溶液。例如，可藉由下述方法來製備無菌注射溶液：在適當的溶劑中摻入必需劑量的本發明的抗體，以及任選地，同時摻入其他期望的成分(包括但不限於，pH調節劑，表面活性劑，佐劑，離子強度增強劑，等滲劑、防腐劑、稀釋劑，或其任何組合)，隨後過濾除菌。此外，可以將無菌注射溶液製備為無菌凍乾粉劑(例如，藉由真空乾燥或冷凍乾燥)以便於儲存和使用。此類無菌凍乾粉劑可在使用前分散於合適的載體中，例如注射用水(WFI)、抑菌性注射用水(BWFI)、氯化鈉溶液（例如0.9% (w/v)NaCl）、葡萄糖溶液（例如5%葡萄糖）、含有表面活性劑的溶液（例如0.01%聚山梨醇20）、pH緩衝溶液（例如磷酸鹽緩衝溶液）、Ringer氏溶液及其任意組合。The antibody or antigen-binding fragment thereof, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or host cell (such as T cell) expressing the chimeric antigen receptor of the present invention can be Formulated into any dosage form known in the medical field, for example, tablets, pills, suspensions, emulsions, solutions, gels, capsules, powders, granules, elixirs, lozenges, suppositories, injections (including injections) Liquid, sterile powder for injection and concentrated solution for injection), inhalation, spray, etc. The preferred dosage form depends on the intended mode of administration and therapeutic use. The pharmaceutical composition of the present invention should be sterile and stable under production and storage conditions. A preferred dosage form is injection. Such injection may be a sterile injection solution. For example, a sterile injectable solution can be prepared by the following method: incorporating the necessary dose of the antibody of the present invention in a suitable solvent, and optionally, at the same time incorporating other desired ingredients (including but not limited to, pH adjusting agent). , Surfactants, adjuvants, ionic strength enhancers, isotonic agents, preservatives, diluents, or any combination thereof), followed by filtration and sterilization. In addition, the sterile injection solution can be prepared as a sterile lyophilized powder (for example, by vacuum drying or freeze drying) for storage and use. Such sterile lyophilized powder can be dispersed in a suitable carrier before use, such as water for injection (WFI), bacteriostatic water for injection (BWFI), sodium chloride solution (for example, 0.9% (w/v) NaCl), glucose Solution (for example, 5% glucose), solution containing surfactant (for example, 0.01% polysorbate 20), pH buffer solution (for example, phosphate buffer solution), Ringer's solution, and any combination thereof.

此外，本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如T細胞）可以以單位劑量形式存在於藥物組合物中，以便於施用。In addition, the antibody or antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or host cell expressing the chimeric antigen receptor (such as T cell) of the present invention ) Can be present in the pharmaceutical composition in a unit dosage form for ease of administration.

本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如T細胞）可以藉由本領域已知的任何合適的方法來施用，包括但不限於，口服、口腔、舌下、眼球、局部、腸胃外、直腸、葉鞘內、內胞漿網槽內、腹股溝、膀胱內、局部（如，粉劑、藥膏或滴劑），或鼻腔途徑。但是，對於許多治療用途而言，較佳的給藥途徑/方式是胃腸外給藥（例如靜脈注射或推注，皮下注射，腹膜內注射，肌內注射）。技術人員應理解，給藥途徑和/或方式將根據預期目的而發生變化。在一個較佳的實施方案中，本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如T細胞）藉由靜脈注射或推注給予。The antibody or antigen-binding fragment thereof, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or host cell (such as T cell) expressing the chimeric antigen receptor of the present invention can be Administration by any suitable method known in the art, including, but not limited to, oral, oral, sublingual, ocular, topical, parenteral, rectal, intralobular, intracytoplasmic reticulum, groin, intravesical, topical (Eg, powder, ointment or drops), or nasal route. However, for many therapeutic purposes, the preferred route/mode of administration is parenteral administration (for example, intravenous injection or bolus injection, subcutaneous injection, intraperitoneal injection, intramuscular injection). The skilled person should understand that the route and/or manner of administration will vary according to the intended purpose. In a preferred embodiment, the antibody of the present invention or its antigen-binding fragment, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimeric antigen receptor or expression of the chimeric antigen receptor The host cells (such as T cells) are given by intravenous injection or bolus injection.

本發明的藥物組合物可以包括“治療有效量”或“預防有效量”的本發明的抗體或其抗原結合片段、雙特異性或多特異性分子、免疫綴合物、藥物組合物、嵌合抗原受體或表達該嵌合抗原受體的宿主細胞（例如T細胞）。“預防有效量”是指，足以預防，阻止，或延遲疾病的發生的量。“治療有效量”是指，足以治癒或至少部分阻止已患有疾病的患者的疾病和其併發症的量。本發明的抗體或其抗原結合片段的治療有效量可根據如下因素發生變化：待治療的疾病的嚴重度、患者自己的免疫系統的總體狀態、患者的一般情況例如年齡，體重和性別，藥物的施用方式，以及同時施用的其他治療等等。The pharmaceutical composition of the present invention may include a "therapeutically effective amount" or "prophylactically effective amount" of the antibody or antigen-binding fragment thereof, bispecific or multispecific molecule, immunoconjugate, pharmaceutical composition, chimera of the present invention An antigen receptor or a host cell (such as a T cell) expressing the chimeric antigen receptor. "Prophylactically effective amount" refers to an amount sufficient to prevent, prevent, or delay the occurrence of a disease. "Therapeutically effective amount" refers to an amount sufficient to cure or at least partially prevent the disease and its complications in patients who have already suffered from the disease. The therapeutically effective amount of the antibody or antigen-binding fragment thereof of the present invention may vary according to the following factors: the severity of the disease to be treated, the overall state of the patient’s own immune system, the patient’s general conditions such as age, weight and gender, and the drug’s Mode of administration, and other treatments administered at the same time, and so on.

在本發明中，可調整給藥方案以獲得最佳目的反應（例如治療或預防反應）。例如，可以單次給藥，可以在一段時間內多次給藥，或者可以隨治療情況的緊急程度按比例減少或增加劑量。In the present invention, the dosage regimen can be adjusted to obtain the best objective response (for example, therapeutic or preventive response). For example, it can be administered in a single dose, it can be administered multiple times over a period of time, or the dose can be reduced or increased proportionally to the urgency of the treatment situation.

在本發明中，該受試者可以為哺乳動物，例如人。檢測方法和試劑盒In the present invention, the subject may be a mammal, such as a human. Detection method and kit

本發明的抗體或其抗原結合片段能夠特異性結合CLDN18.2，從而可用於檢測CLDN18.2在樣品中的存在或其水準。The antibody or antigen-binding fragment thereof of the present invention can specifically bind to CLDN 18.2, and thus can be used to detect the presence or level of CLDN 18.2 in a sample.

因此，在另一個方面，本發明提供了一種試劑盒，其包括本發明的抗體或其抗原結合片段。在某些較佳的實施方案中，本發明的抗體或其抗原結合片段帶有可檢測的標記。在一個較佳的實施方案中，該試劑盒還包括第二抗體，其特異性識別本發明的抗體或其抗原結合片段。較佳地，該第二抗體還包括可檢測的標記。Therefore, in another aspect, the present invention provides a kit comprising the antibody of the present invention or an antigen-binding fragment thereof. In some preferred embodiments, the antibody or antigen-binding fragment thereof of the present invention bears a detectable label. In a preferred embodiment, the kit further includes a second antibody, which specifically recognizes the antibody of the present invention or an antigen-binding fragment thereof. Preferably, the second antibody also includes a detectable label.

在本發明中，該可檢測的標記可以是可藉由螢光、光譜、光化學、生物化學、免疫學、電學、光學或化學手段檢測的任何物質。特別佳的是，此類標記能夠適用於免疫學檢測（例如，酶聯免疫測定法、放射免疫測定法、螢光免疫測定法、化學發光免疫測定法等）。這類標記是本領域熟知的，包括但不限於，酶（例如，辣根過氧化物酶、鹼性磷酸酶、β-半乳糖苷酶、脲酶、葡萄糖氧化酶，等）、放射性核素（例如，³ H、¹²⁵ I、³⁵ S、¹⁴ C或³² P）、螢光染料（例如，異硫氰酸螢光素（FITC）、螢光素、異硫氰酸四甲基羅丹明（TRITC）、藻紅蛋白（PE）、德克薩斯紅、羅丹明、量子點或花菁染料衍生物（例如Cy7、Alexa 750））、發光物質（例如化學發光物質，如吖啶酯類化合物）、磁珠（例如，Dynabeads^® ）、測熱標記物例如膠體金或有色玻璃或塑膠（例如，聚苯乙烯、聚丙烯、乳膠，等）珠、以及用於結合上述標記物修飾的親和素（例如，鏈黴親和素）的生物素。教導該標記物的使用的專利包括，但不限於，美國專利3,817,837；3,850,752；3,939,350；3,996,345；4,277,437；4,275,149；及4,366,241（全部藉由引用併入本文）。本發明中涵蓋的標記物可藉由本領域已知的方法檢測。例如，放射性標記可使用攝影膠片或閃爍計算器檢測，螢光標記物可使用光檢測器檢測，以檢測發射的光。酶標記物一般藉由給酶提供底物及檢測藉由酶對底物的作用產生的反應產物來檢測，及測熱標記物藉由簡單視覺化著色標記物來檢測。在某些實施方案中，可藉由不同長度的接頭將如上所述的可檢測的標記連接至本發明的抗體或其抗原結合片段，以降低潛在的位阻。In the present invention, the detectable label can be any substance that can be detected by fluorescence, spectroscopy, photochemistry, biochemistry, immunology, electrical, optical or chemical means. It is particularly preferable that such a label can be applied to immunological detection (for example, enzyme-linked immunoassay, radioimmunoassay, fluoroimmunoassay, chemiluminescence immunoassay, etc.). Such labels are well known in the art and include, but are not limited to, enzymes (for example, horseradish peroxidase, alkaline phosphatase, β-galactosidase, urease, glucose oxidase, etc.), radionuclides ( For example, ³ H, ¹²⁵ I, ³⁵ S, ¹⁴ C or ³² P), fluorescent dyes (for example, fluorescein isothiocyanate (FITC), luciferin, tetramethylrhodamine isothiocyanate (TRITC) ), phycoerythrin (PE), Texas red, rhodamine, quantum dots or cyanine dye derivatives (such as Cy7, Alexa 750)), luminescent substances (such as chemiluminescent substances, such as acridine ester compounds) , Magnetic beads (for example, Dynabeads ^® ), calorimetric markers such as colloidal gold or colored glass or plastic (for example, polystyrene, polypropylene, latex, etc.) beads, and avidin ( For example, streptavidin) biotin. Patents teaching the use of this marker include, but are not limited to, US Patent Nos. 3,817,837; 3,850,752; 3,939,350; 3,996,345; 4,277,437; 4,275,149; and 4,366,241 (all incorporated herein by reference). The markers covered by the present invention can be detected by methods known in the art. For example, a radioactive label can be detected using photographic film or a scintillation calculator, and a fluorescent label can be detected using a light detector to detect the emitted light. Enzyme markers are generally detected by providing a substrate for the enzyme and detecting reaction products produced by the action of the enzyme on the substrate, and calorimetric markers are detected by simply visualizing colored markers. In some embodiments, the detectable label as described above can be connected to the antibody or antigen-binding fragment thereof of the present invention by linkers of different lengths to reduce potential steric hindrance.

在另一個方面，本發明提供了檢測CLDN18.2在樣品中的存在或其量的方法，其包括以下步驟： (1) 將該樣品與本發明的抗體或其抗原結合片段接觸； (2) 檢測該抗體或其抗原結合片段與CLDN18.2之間複合物的形成或檢測該複合物的量。In another aspect, the present invention provides a method for detecting the presence or amount of CLDN 18.2 in a sample, which includes the following steps: (1) Contact the sample with the antibody or antigen-binding fragment thereof of the present invention; (2) Detect the formation of the complex between the antibody or its antigen-binding fragment and CLDN18.2 or detect the amount of the complex.

該複合物的形成表明存在CLDN18.2或表達CLDN18.2的細胞。The formation of this complex indicates the presence of CLDN18.2 or cells expressing CLDN18.2.

在某些較佳的實施方案中，該樣品是細胞樣品，即包含細胞(例如腫瘤細胞)的樣品。在此類實施方案中，較佳地，該複合物是由該抗體、抗原結合片段或綴合物與該樣品中的細胞所表達的CLDN18.2之間形成的。In certain preferred embodiments, the sample is a cell sample, that is, a sample containing cells (e.g., tumor cells). In such embodiments, preferably, the complex is formed between the antibody, antigen-binding fragment or conjugate and CLDN 18.2 expressed by the cells in the sample.

在一個較佳的實施方案中，本發明的抗體或其抗原結合片段還帶有可檢測的標記。在另一個較佳的實施方案中，在步驟(2)中，使用帶有可檢測的標記的試劑來檢測本發明的抗體或其抗原結合片段。In a preferred embodiment, the antibody or antigen-binding fragment thereof of the present invention also bears a detectable label. In another preferred embodiment, in step (2), a reagent with a detectable label is used to detect the antibody or antigen-binding fragment thereof of the present invention.

該方法可以用於診斷目的，或者非診斷目的（例如，該樣品是細胞樣品，而非來自患者的樣品）。在某些較佳的實施方案中，該CLDN18.2是人CLDN18.2。The method can be used for diagnostic or non-diagnostic purposes (for example, the sample is a cell sample, not a sample from a patient). In certain preferred embodiments, the CLDN 18.2 is human CLDN 18.2.

在另一個方面，提供了本發明的抗體或其抗原結合片段在製備試劑盒中的用途，該試劑盒用於檢測CLDN18.2在樣品中的存在或其量。在某些較佳的實施方案中，該CLDN18.2是人CLDN18.2。In another aspect, there is provided the use of the antibody or antigen-binding fragment thereof of the present invention in the preparation of a kit for detecting the presence or amount of CLDN 18.2 in a sample. In certain preferred embodiments, the CLDN 18.2 is human CLDN 18.2.

在另一個方面，本發明提供了一種用於檢測腫瘤是否能夠藉由靶向CLDN18.2的抗腫瘤療法來治療的方法，其包括以下步驟： (1) 將含有該腫瘤細胞的樣品與本發明的抗體或其抗原結合片段接觸； (2) 檢測該抗體或其抗原結合片段與CLDN18.2之間複合物的形成。In another aspect, the present invention provides a method for detecting whether a tumor can be treated by an anti-tumor therapy targeting CLDN 18.2, which includes the following steps: (1) Contacting the sample containing the tumor cells with the antibody or antigen-binding fragment thereof of the present invention; (2) Detect the formation of the complex between the antibody or its antigen-binding fragment and CLDN18.2.

在某些較佳的實施方案中，該複合物是該抗體或其抗原結合片段與該樣品中的腫瘤細胞所表達的CLDN18.2之間形成的。In certain preferred embodiments, the complex is formed between the antibody or antigen-binding fragment thereof and CLDN 18.2 expressed by tumor cells in the sample.

在某些較佳的實施方案中，該樣品來自受試者，該受試者患有腫瘤、懷疑患有腫瘤或具有患有腫瘤風險。在某些較佳的實施方案中，該樣品來自在組織或器官未患癌症時其中的細胞基本不表達CLDN18.2的組織或器官。在某些較佳的實施方案中，該組織選自胃組織、肺組織、食道組織、胰腺組織或乳腺組織，並且該組織任選地已被診斷為被癌症影響，例如藉由對該組織或器官的細胞進行目視檢查或培養測試被診斷。在某些較佳的實施方案中，該組織是除胃組織以外的組織。在某些較佳的實施方案中，該組織是肺組織、食道組織、胰腺組織或乳腺組織。在此類實施方案中，當存在CLDN18.2或表達CLDN18.2的細胞，和/或與參照水準相比(例如與無腫瘤疾病的患者相比)，CLDN18.2或表達CLDN18.2的細胞的量增加時，表明該受試者適於靶向CLDN18.2的抗腫瘤療法。In certain preferred embodiments, the sample is from a subject who has a tumor, is suspected of having a tumor, or is at risk of having a tumor. In some preferred embodiments, the sample is from a tissue or organ whose cells do not substantially express CLDN 18.2 when the tissue or organ does not suffer from cancer. In certain preferred embodiments, the tissue is selected from stomach tissue, lung tissue, esophagus tissue, pancreas tissue, or breast tissue, and the tissue has optionally been diagnosed as being affected by cancer, for example, by The organ's cells are diagnosed by visual inspection or culture test. In certain preferred embodiments, the tissue is a tissue other than stomach tissue. In certain preferred embodiments, the tissue is lung tissue, esophagus tissue, pancreas tissue, or breast tissue. In such embodiments, when CLDN18.2 or cells expressing CLDN18.2 are present, and/or compared to a reference level (e.g., compared to patients without tumor disease), CLDN18.2 or cells expressing CLDN18.2 The increase in the amount indicates that the subject is suitable for anti-tumor therapy targeting CLDN 18.2.

在某些較佳的實施方案中，該CLDN18.2是人CLDN18.2。In certain preferred embodiments, the CLDN 18.2 is human CLDN 18.2.

在某些較佳的實施方案中，該腫瘤選自食道癌、胰腺癌、支氣管癌、非小細胞肺癌、乳腺癌、耳鼻喉(ENT)癌、卵巢癌、結腸癌、肝癌、頭頸癌、膽囊癌及其轉移癌（例如，胃癌轉移，如Krukenberg瘤、腹膜轉移或淋巴結轉移。In certain preferred embodiments, the tumor is selected from the group consisting of esophageal cancer, pancreatic cancer, bronchial cancer, non-small cell lung cancer, breast cancer, ear nose and throat (ENT) cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer, gallbladder Cancer and its metastases (for example, gastric cancer metastasis, such as Krukenberg tumor, peritoneal metastasis, or lymph node metastasis.

在另一個方面，提供了本發明的抗體或其抗原結合片段在製備試劑盒中的用途，該試劑盒用於檢測腫瘤是否能夠藉由靶向CLDN18.2的抗腫瘤療法來治療。In another aspect, there is provided the use of the antibody or antigen-binding fragment thereof of the present invention in the preparation of a kit for detecting whether a tumor can be treated by anti-tumor therapy targeting CLDN 18.2.

在某些較佳的實施方案中，該抗體或其抗原結合片段帶有可檢測的標記。In certain preferred embodiments, the antibody or antigen-binding fragment thereof bears a detectable label.

在某些較佳的實施方案中，該腫瘤選自食道癌、胰腺癌、支氣管癌、非小細胞肺癌、乳腺癌、耳鼻喉(ENT)癌、卵巢癌、結腸癌、肝癌、頭頸癌、膽囊癌及其轉移癌（例如，胃癌轉移，如Krukenberg瘤、腹膜轉移或淋巴結轉移。術語定義In certain preferred embodiments, the tumor is selected from the group consisting of esophageal cancer, pancreatic cancer, bronchial cancer, non-small cell lung cancer, breast cancer, ear nose and throat (ENT) cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer, gallbladder Cancer and its metastases (for example, gastric cancer metastasis, such as Krukenberg tumor, peritoneal metastasis, or lymph node metastasis. Definition of Terms

在本發明中，除非另有說明，否則本文中使用的科學和技術名詞具有本領域技術人員所通常理解的含義。並且，本文中所用的細胞培養、生物化學、核酸化學、免疫學實驗室等操作步驟均為相應領域內廣泛使用的常規步驟。同時，為了更好地理解本發明，下面提供相關術語的定義和解釋。In the present invention, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. In addition, the cell culture, biochemistry, nucleic acid chemistry, immunology laboratory and other operating steps used in this article are all routine steps widely used in the corresponding fields. At the same time, in order to better understand the present invention, definitions and explanations of related terms are provided below.

如本文中所使用的，術語“CLDN18（Claudin 18，密蛋白18）”具有本領域技術人員通常理解的含義，其屬於密蛋白（Claudin）家族，是上皮與內皮的緊密連接內的跨膜蛋白，並且以兩種剪接變體CLDN18.1和CLDN18.2的形式存在。CLDN18.1及CLDN18.2的序列均是本領域公知的，可分別參見NCBI資料庫登錄號NP_057453.1和NP_001002026.1。As used herein, the term “CLDN18 (Claudin 18, claudin 18)” has the meaning commonly understood by those skilled in the art. It belongs to the Claudin family and is a transmembrane protein within the tight junction of epithelium and endothelium. , And exist in the form of two splice variants CLDN18.1 and CLDN18.2. The sequences of CLDN18.1 and CLDN18.2 are well known in the art, and can be referred to NCBI database accession numbers NP_057453.1 and NP_001002026.1, respectively.

如本文中所使用的，術語“抗體”是指，通常由兩對多肽鏈（每對具有一條輕鏈(LC)和一條重鏈(HC)）組成的免疫球蛋白分子。抗體輕鏈可分類為κ（kappa）和λ（lambda）輕鏈。重鏈可分類為μ、δ、γ、α或ε，並且分別將抗體的同種型定義為IgM、IgD、IgG、IgA和IgE。在輕鏈和重鏈內，可變區和恆定區藉由大約12或更複數氨基酸的“J”區連接，重鏈還包含大約3個或更複數氨基酸的“D”區。各重鏈由重鏈可變區(VH)和重鏈恆定區(CH)組成。重鏈恆定區由3個結構域(CH1、CH2和CH3)組成。各輕鏈由輕鏈可變區(VL)和輕鏈恆定區(CL)組成。輕鏈恆定區由一個結構域CL組成。恆定結構域不直接參與抗體與抗原的結合，但展現出多種效應子功能，如可介導免疫球蛋白與宿主組織或因數，包括免疫系統的各種細胞(例如，效應細胞)和經典補體系統的第一組分(C1q)的結合。VH和VL區還可被細分為具有高變性的區域(稱為互補決定區(CDR))，其間散佈有較保守的稱為構架區(FR)的區域。各V_H 和V_L 由按下列順序：FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4從氨基末端至羧基末端排列的3個CDR和4個FR組成。各重鏈/輕鏈對的可變區(VH和VL)分別形成抗原結合部位。氨基酸在各區域或結構域的分配可遵循Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991))，或Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia等人 (1989) Nature 342:878-883的定義。As used herein, the term "antibody" refers to an immunoglobulin molecule usually composed of two pairs of polypeptide chains (each pair has a light chain (LC) and a heavy chain (HC)). Antibody light chains can be classified into kappa (kappa) and lambda (lambda) light chains. Heavy chains can be classified as mu, delta, gamma, alpha, or epsilon, and the isotype of the antibody is defined as IgM, IgD, IgG, IgA, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are connected by a "J" region of about 12 or more amino acids, and the heavy chain also includes a "D" region of about 3 or more amino acids. Each heavy chain is composed of a heavy chain variable region (VH) and a heavy chain constant region (CH). The heavy chain constant region is composed of 3 domains (CH1, CH2, and CH3). Each light chain is composed of a light chain variable region (VL) and a light chain constant region (CL). The constant region of the light chain consists of a domain CL. The constant domains are not directly involved in the binding of antibodies and antigens, but exhibit a variety of effector functions, such as mediating immunoglobulins and host tissues or factors, including various cells of the immune system (for example, effector cells) and the classical complement system. Combination of the first component (C1q). The VH and VL regions can also be subdivided into regions with hyperdenaturation (called complementarity determining regions (CDR)), interspersed with more conservative regions called framework regions (FR). Each V _H and V _L, the following order: FR1, CDR1, FR2, CDR2 , FR3, CDR3, FR4 from the amino terminus to the carboxy terminus arranged three four FR and CDR components. The variable regions (VH and VL) of each heavy chain/light chain pair respectively form an antigen binding site. The assignment of amino acids in each region or domain can follow Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, Md. (1987 and 1991)), or Chothia & Lesk (1987) J. Mol. Biol. 196 :901-917; Definition of Chothia et al. (1989) Nature 342:878-883.

如本文中所使用的，術語“互補決定區”或“CDR”是指抗體可變區中負責抗原結合的氨基酸殘基。在重鏈和輕鏈的可變區中各含有三個CDR，命名為CDR1、CDR2和CDR3。這些CDR的精確邊界可根據本領域已知的各種編號系統進行定義，例如可按照Kabat編號系統（Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md., 1991）、Chothia編號系統（Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia等人 (1989) Nature 342:878-883）或IMGT編號系統（Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003）中的定義。對於給定的抗體，本領域技術人員將容易地鑒別各編號系統所定義的CDR。並且，不同編號系統之間的對應關係是本領域技術人員熟知的（例如，可參見Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003）。As used herein, the term "complementarity determining region" or "CDR" refers to the amino acid residues in the variable region of an antibody that are responsible for antigen binding. Each of the variable regions of the heavy chain and the light chain contains three CDRs, named CDR1, CDR2, and CDR3. The precise boundaries of these CDRs can be defined according to various numbering systems known in the art, for example, according to the Kabat numbering system (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda , Md., 1991), Chothia numbering system (Chothia & Lesk (1987) J. Mol. Biol. 196:901-917; Chothia et al. (1989) Nature 342:878-883) or IMGT numbering system (Lefranc et al ., Dev. Comparat. Immunol. 27:55-77, 2003). For a given antibody, those skilled in the art will easily identify the CDRs defined by each numbering system. Moreover, the correspondence between different numbering systems is well known to those skilled in the art (for example, see Lefranc et al., Dev. Comparat. Immunol. 27:55-77, 2003).

在本發明中，本發明的抗體或其抗原結合片段含有的CDR可根據本領域已知的各種編號系統確定。在某些實施方案中，本發明的抗體或其抗原結合片段含有的CDR較佳地藉由Kabat、Chothia或IMGT編號系統確定。在某些實施方案中，本發明的抗體或其抗原結合片段含有的CDR較佳地藉由Kabat編號系統確定。In the present invention, the CDR contained in the antibody or antigen-binding fragment thereof of the present invention can be determined according to various numbering systems known in the art. In certain embodiments, the CDR contained in the antibody or antigen-binding fragment thereof of the present invention is preferably determined by the Kabat, Chothia or IMGT numbering system. In certain embodiments, the CDR contained in the antibody or antigen-binding fragment thereof of the present invention is preferably determined by the Kabat numbering system.

如本文中所使用的，術語“構架區（framework region）”或“FR”殘基是指，抗體可變區中除了如上定義的CDR殘基以外的那些氨基酸殘基。As used herein, the term "framework region" or "FR" residues refers to those amino acid residues in the variable region of an antibody other than the CDR residues as defined above.

術語“抗體”不受任何特定的產生抗體的方法限制。例如，其包括，重組抗體、單殖株抗體和多殖株抗體。抗體可以是不同同種型的抗體，例如，IgG (例如，IgG1，IgG2，IgG3或IgG4亞型)，IgA1，IgA2，IgD，IgE或IgM抗體。The term "antibody" is not limited by any specific method of producing antibodies. For example, it includes recombinant antibodies, monoclonal antibodies, and polyclonal antibodies. The antibodies may be antibodies of different isotypes, for example, IgG (eg, IgG1, IgG2, IgG3 or IgG4 subtype), IgA1, IgA2, IgD, IgE or IgM antibodies.

如本文中所使用的，術語抗體的“抗原結合片段”是指包含全長抗體的片段的多肽，其保持特異性結合全長抗體所結合的相同抗原的能力，和/或與全長抗體競爭對抗原的特異性結合，其也被稱為“抗原結合部分”。通常參見，Fundamental Immunology, Ch. 7 (Paul, W., ed., 第2版，Raven Press, N.Y. (1989)，其以其全文藉由引用合併入本文，用於所有目的。可藉由重組DNA技術或藉由完整抗體的酶促或化學斷裂產生抗體的抗原結合片段。抗原結合片段的非限制性實例包括Fab、Fab’、F(ab’)₂ 、Fd、Fv、互補決定區(CDR)片段、scFv、雙抗體（diabody）、單域抗體（single domain antibody）、嵌合抗體、線性抗體（linear antibody）、奈米抗體（技術來自Domantis）、、probody和這樣的多肽，其包含足以賦予多肽特異性抗原結合能力的抗體的至少一部分。工程改造的抗體變體綜述於Holliger等, 2005; Nat Biotechnol, 23: 1126-1136中。As used herein, the term “antigen-binding fragment” of an antibody refers to a polypeptide comprising a fragment of a full-length antibody that retains the ability to specifically bind to the same antigen that the full-length antibody binds, and/or competes with the full-length antibody for antigen Specific binding, which is also called "antigen binding portion". See generally, Fundamental Immunology, Ch. 7 (Paul, W., ed., 2nd edition, Raven Press, NY (1989), which is incorporated herein by reference in its entirety for all purposes. Can be reorganized DNA technology or by enzymatic or chemical fragmentation of intact antibodies to produce antigen-binding fragments of antibodies. Non-limiting examples of antigen-binding fragments include Fab, Fab', F(ab') ₂ , Fd, Fv, complementarity determining region (CDR) ) Fragment, scFv, diabody, single domain antibody, chimeric antibody, linear antibody, nano antibody (technology from Domantis), probody and such polypeptides, which contain sufficient At least a part of an antibody that confers specific antigen binding ability to a polypeptide. Engineered antibody variants are reviewed in Holliger et al., 2005; Nat Biotechnol, 23: 1126-1136.

如本文中所使用的，術語“全長抗體”意指，由兩條“全長重鏈”和兩條“全長輕鏈”組成的抗體。其中，“全長重鏈”是指這樣的多肽鏈，其在N端到C端的方向上由重鏈可變區（VH）、重鏈恆定區CH1結構域、鉸鏈區（HR）、重鏈恆定區CH2結構域、重鏈恆定區CH3結構域組成；並且，當所述全長抗體為IgE同種型時，任選地還包括重鏈恆定區CH4結構域。較佳地，“全長重鏈”是在N端到C端方向上由VH、CH1、HR、CH2和CH3組成的多肽鏈。“全長輕鏈”是在N端到C端方向上由輕鏈可變區(VL)和輕鏈恆定區（CL）組成的多肽鏈。兩對全長抗體鏈藉由在CL和CH1之間的二硫鍵和兩條全長重鏈的HR之間的二硫鍵連接在一起。本發明的全長抗體可以來自單一物種，例如人；也可以是嵌合抗體或人源化抗體。本發明的全長抗體包含分別由VH和VL對形成的兩個抗原結合部位，這兩個抗原結合部位特異性識別/結合相同的抗原。As used herein, the term "full-length antibody" means an antibody composed of two "full-length heavy chains" and two "full-length light chains." Among them, "full-length heavy chain" refers to a polypeptide chain that consists of a heavy chain variable region (VH), a heavy chain constant region CH1 domain, a hinge region (HR), and a constant heavy chain in the N-terminal to C-terminal direction. The region consists of a CH2 domain and a heavy chain constant region CH3 domain; and, when the full-length antibody is of an IgE isotype, it optionally also includes a heavy chain constant region CH4 domain. Preferably, the "full-length heavy chain" is a polypeptide chain composed of VH, CH1, HR, CH2, and CH3 in the N-terminal to C-terminal direction. A "full-length light chain" is a polypeptide chain composed of a light chain variable region (VL) and a light chain constant region (CL) in the N-terminal to C-terminal direction. The two pairs of full-length antibody chains are linked together by a disulfide bond between CL and CH1 and a disulfide bond between the HR of the two full-length heavy chains. The full-length antibody of the present invention can be from a single species, such as human; it can also be a chimeric antibody or a humanized antibody. The full-length antibody of the present invention includes two antigen binding sites formed by a pair of VH and VL respectively, and the two antigen binding sites specifically recognize/bind the same antigen.

如本文中所使用的，術語“Fd”意指由VH和CH1結構域組成的抗體片段；術語“dAb片段”意指由VH結構域組成的抗體片段（Ward 等人, Nature 341:544 546 (1989)）；術語“Fab片段”意指由VL、VH、CL和CH1結構域組成的抗體片段；術語“F(ab’)₂ 片段”意指包含藉由鉸鏈區上的二硫橋連接的兩個Fab片段的抗體片段；術語“Fab’片段”意指還原連接F(ab’)₂ 片段中兩個重鏈片段的二硫鍵後所獲片段，由一條完整的輕鏈和重鏈的Fd片段（由VH和CH1結構域組成）組成。As used herein, the term "Fd" means an antibody fragment composed of VH and CH1 domains; the term "dAb fragment" means an antibody fragment composed of VH domains (Ward et al., Nature 341:544 546 ( 1989)); the term "Fab fragment" means an antibody fragment composed of VL, VH, CL and CH1 domains; the term "F(ab') ₂ fragment" means a fragment comprising a disulfide bridge connected by a hinge region The antibody fragment of two Fab fragments; the term "Fab'fragment" means the _{fragment obtained by reducing the disulfide bond connecting the two heavy chain fragments in the F(ab') 2} fragment, consisting of a complete light chain and heavy chain Fd fragment (consisting of VH and CH1 domains).

如本文中所使用的，術語“Fv”意指由抗體的單臂的VL和VH結構域組成的抗體片段。Fv片段通常被認為是，能形成完整的抗原結合位點的最小抗體片段。一般認為，六個CDR賦予抗體的抗原結合特異性。然而，即便是一個可變區（例如Fd片段，其僅僅含有三個對抗原特異的CDR）也能夠識別並結合抗原，儘管其親和力可能低於完整的結合位點。As used herein, the term "Fv" means an antibody fragment composed of the VL and VH domains of a single arm of an antibody. Fv fragments are generally considered to be the smallest antibody fragments that can form a complete antigen binding site. It is generally believed that the six CDRs confer antigen-binding specificity to an antibody. However, even a variable region (such as an Fd fragment, which contains only three antigen-specific CDRs) can recognize and bind antigen, although its affinity may be lower than the complete binding site.

如本文中所使用的，術語“Fc”意指，由抗體的第一重鏈的第二、第三恆定區與第二重鏈的第二、第三恆定區經二硫鍵結合而形成的抗體片段。抗體的Fc片段具有多種不同的功能，但不參與抗原的結合。As used herein, the term "Fc" means a disulfide bond formed by the second and third constant regions of the first heavy chain of an antibody and the second and third constant regions of the second heavy chain. Antibody fragments. The Fc fragment of an antibody has many different functions, but does not participate in antigen binding.

如本文中所使用的，術語“scFv”是指，包含VL和VH結構域的單個多肽鏈，其中所述VL和VH藉由接頭（linker）相連（參見，例如；Bird等人， Science 242:423-426 (1988)；Huston等人，Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988)；和Pluckthun，The Pharmacology of Monoclonal Antibodies，第113卷， Roseburg 和Moore 編，Springer-Verlag，紐約，第269-315頁(1994)）。此類scFv分子可具有一般結構：NH₂ -VL-接頭-VH-COOH或NH₂ -VH-接頭-VL-COOH。合適的先前技術接頭由重複的GGGGS氨基酸序列或其變體組成。例如，可使用具有氨基酸序列(GGGGS)₄ 的接頭，但也可使用其變體（Holliger等人(1993)，Proc. Natl. Acad. Sci. USA 90: 6444-6448）。可用於本發明的其他接頭由Alfthan等人(1995)，Protein Eng. 8:725-731，Choi等人(2001)，Eur. J. Immunol. 31: 94-106，Hu等人(1996)，Cancer Res. 56:3055-3061，Kipriyanov等人(1999)，J. Mol. Biol. 293:41-56和Roovers等人(2001)，Cancer Immunol.描述。在一些情況下，scFv的VH與VL之間還可以存在二硫鍵。As used herein, the term "scFv" refers to a single polypeptide chain comprising VL and VH domains, where the VL and VH are connected by a linker (see, for example; Bird et al., Science 242: 423-426 (1988); Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Pluckthun, The Pharmacology of Monoclonal Antibodies, Volume 113, Roseburg and Moore eds, Springer-Verlag , New York, pp. 269-315 (1994)). Such scFv molecules may have the general structure: NH ₂ -VL-linker-VH-COOH or NH _{2 -VH-linker-VL-COOH.} Suitable prior art linkers consist of repeated GGGGS amino acid sequences or variants thereof. For example, a linker having an amino acid sequence (GGGGS) ₄ can be used, but variants thereof can also be used (Holliger et al. (1993), Proc. Natl. Acad. Sci. USA 90: 6444-6448). Other linkers that can be used in the present invention are described by Alfthan et al. (1995), Protein Eng. 8:725-731, Choi et al. (2001), Eur. J. Immunol. 31: 94-106, Hu et al. (1996), Cancer Res. 56:3055-3061, Kipriyanov et al. (1999), J. Mol. Biol. 293:41-56 and Roovers et al. (2001), Cancer Immunol. In some cases, there may also be disulfide bonds between the VH and VL of the scFv.

如本文中所使用的，術語“雙抗體”意指，其VH和VL結構域在單個多肽鏈上表達，但使用太短的連接體以致不允許在相同鏈的兩個結構域之間配對，從而迫使結構域與另一條鏈的互補結構域配對並且產生兩個抗原結合部位（參見，例如, Holliger P.等人, Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993)，和Poljak R. J.等人, Structure 2:1121-1123 (1994)）。As used herein, the term "diabody" means that its VH and VL domains are expressed on a single polypeptide chain, but a linker that is too short to allow pairing between the two domains of the same chain, Thereby forcing the domain to pair with the complementary domain of the other chain and create two antigen binding sites (see, for example, Holliger P. et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993), and Poljak RJ et al., Structure 2:1121-1123 (1994)).

如本文中所使用的，術語“單域抗體（single-domain antibody, sdAb）”具有本領域技術人員通常理解的含義，其是指由單個單體可變抗體結構域（例如單個重鏈可變區）所組成的抗體片段，其保持特異性結合全長抗體所結合的相同抗原的能力。單域抗體也稱為奈米抗體（nanobody）。As used herein, the term "single-domain antibody (sdAb)" has the meaning commonly understood by those skilled in the art, which refers to a single monomer variable antibody domain (for example, a single heavy chain variable antibody). Region), which retains the ability to specifically bind to the same antigen bound by the full-length antibody. Single domain antibodies are also called nanobodies.

如本文中所使用的，術語“probody”具有本領域技術人員通常理解的含義，其是指一種經掩蔽的抗體，其在健康組織中保持惰性，但在疾病環境中特異性活化(例如，經由疾病環境中富集或特有的蛋白酶的蛋白酶裂解)。其詳細教導可參見，例如Desnoyers等人，Sci.Transl.Med.,5:207ra144,2013。可對本文所描述的任何抗體或其抗原結合部分使用類似掩蔽技術。As used herein, the term "probody" has the meaning commonly understood by those skilled in the art, which refers to a masked antibody that remains inert in healthy tissues but is specifically activated in the disease environment (e.g., via Protease cleavage of proteases that are enriched or unique in the disease environment). For detailed teaching, see, for example, Desnoyers et al., Sci. Transl. Med., 5:207ra144, 2013. Similar masking techniques can be used on any antibody or antigen binding portion thereof described herein.

上述各個抗體片段均保持了特異性結合全長抗體所結合的相同抗原的能力，和/或與全長抗體競爭對抗原的特異性結合。Each of the aforementioned antibody fragments retains the ability to specifically bind to the same antigen that the full-length antibody binds, and/or competes with the full-length antibody for specific binding to the antigen.

可使用本領域技術人員已知的常規技術（例如，重組DNA技術或酶促或化學斷裂法）從給定的抗體（例如本發明提供的抗體）獲得抗體的抗原結合片段（例如，上述抗體片段），並且以與用於完整抗體的方式相同的方式就特異性篩選抗體的抗原結合片段。Conventional techniques known to those skilled in the art (for example, recombinant DNA technology or enzymatic or chemical fragmentation) can be used to obtain antigen-binding fragments of antibodies (for example, the above-mentioned antibody fragments) from a given antibody (for example, the antibody provided by the present invention). ), and specifically screen for antigen-binding fragments of antibodies in the same manner as used for intact antibodies.

在本文中，除非上下文明確指出，否則當提及術語“抗體”時，其不僅包括完整抗體，而且包括抗體的抗原結合片段。Herein, unless the context clearly dictates otherwise, when the term "antibody" is referred to, it includes not only intact antibodies but also antigen-binding fragments of antibodies.

如本文中所使用的，術語“單殖株抗體”、“單抗”、“mAb”具有相同的含義且可互換使用可互換，其是指，來自一群高度同源的抗體分子中的一個抗體或抗體的一個片段，也即，除可能自發出現的自然突變外，一群完全相同的抗體分子。單抗對抗原上的單一表位具有高特異性。多殖株抗體是相對於單殖株抗體而言的，其通常包含至少2種或更多種的不同抗體，這些不同的抗體通常識別抗原上的不同表位。此外，修飾語“單殖株”僅表明該抗體的特徵為從高度同源的抗體群中獲得，不能理解為需要藉由任何特定方法來製備該抗體。As used herein, the terms "monoclonal antibody", "monoclonal antibody", and "mAb" have the same meaning and are used interchangeably, which refers to an antibody from a group of highly homologous antibody molecules Or a fragment of an antibody, that is, a group of identical antibody molecules except for natural mutations that may occur spontaneously. The monoclonal antibody has high specificity for a single epitope on the antigen. Polyclonal antibodies are relative to monoclonal antibodies, which usually include at least two or more different antibodies, and these different antibodies usually recognize different epitopes on the antigen. In addition, the modifier "monoclonal strain" only indicates that the antibody is characterized as being obtained from a highly homologous group of antibodies, and cannot be understood as requiring any specific method to prepare the antibody.

本發明的單殖株抗體可以藉由多種技術進行製備，例如雜交瘤技術（參見，例如Kohler等人. Nature, 256:495 ,1975），重組DNA技術（參見，例如美國專利申請4,816,567），或噬菌體抗體庫技術（參見，例如Clackson等. Nature352 ：624-628, 1991，或Marks等. J.Mol.Biol.222 ：581-597, 1991）。The monoclonal antibody of the present invention can be prepared by a variety of techniques, such as hybridoma technology (see, for example, Kohler et al. Nature, 256:495, 1975), recombinant DNA technology (see, for example, U.S. Patent Application 4,816,567), or Phage antibody library technology (see, for example, Clackson et al. Nature352: 624-628, 1991, or Marks et al. J. Mol. Biol. 222: 581-597, 1991).

抗體可藉由公知的技術，例如使用蛋白A或蛋白G的親和層析進行純化。隨後或作為替代，可將特異性抗原（該抗體識別的靶分子）或其抗原表位固定在柱上，並藉由免疫親合層析法來純化免疫特異性抗體。免疫球蛋白的純化可參考例如D.Wilkinson(The Scientist, published by The Scientist, Inc., Philadelphia Pa., Vol.14, No.8(Apr.17,2000), pp.25-28)。Antibodies can be purified by known techniques, such as affinity chromatography using protein A or protein G. Subsequently or as an alternative, the specific antigen (the target molecule recognized by the antibody) or its epitope can be immobilized on the column, and the immunospecific antibody can be purified by immunoaffinity chromatography. For the purification of immunoglobulin, refer to, for example, D. Wilkinson (The Scientist, published by The Scientist, Inc., Philadelphia Pa., Vol. 14, No. 8 (Apr. 17, 2000), pp. 25-28).

如本文中所使用的，術語“嵌合抗體（Chimeric antibody）”是指，這樣的抗體，其輕鏈或/和重鏈的一部分源自一個抗體（其可以源自某一特定物種或屬於某一特定抗體類或亞類），且輕鏈或/和重鏈的另一部分源自另一個抗體（其可以源自相同或不同的物種或屬於相同或不同的抗體類或亞類），但無論如何，其仍保留對目標抗原的結合活性（U.S.P 4,816,567 to Cabilly et al.; Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851 6855 (1984)）。例如，術語“嵌合抗體”可包括這樣的抗體（例如人鼠嵌合抗體），其中抗體的重鏈和輕鏈可變區來自第一抗體（例如鼠源抗體），而抗體的重鏈和輕鏈恆定區來自第二抗體（例如人抗體）。As used herein, the term "chimeric antibody" refers to an antibody whose light chain or/and part of its heavy chain is derived from an antibody (which may be derived from a specific species or belong to a certain species). A specific antibody class or subclass), and another part of the light chain or/and heavy chain is derived from another antibody (which can be derived from the same or different species or belong to the same or different antibody class or subclass), but no matter However, it still retains the binding activity to the target antigen (USP 4,816,567 to Cabilly et al.; Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851 6855 (1984)). For example, the term "chimeric antibody" may include antibodies (e.g., human-mouse chimeric antibodies) in which the heavy and light chain variable regions of the antibody are derived from the first antibody (e.g., murine antibody), and the heavy chain and The light chain constant region is derived from a second antibody (such as a human antibody).

如本文中所使用的，術語“人源化抗體”是指，經基因工程改造的非人源抗體，其氨基酸序列經修飾以提高與人源抗體的序列的同源性。通常而言，人源化抗體的全部或部分CDR區來自於非人源抗體（供體抗體），全部或部分的非CDR區（例如，可變區FR和/或恆定區）來自於人源免疫球蛋白（受體抗體）。人源化抗體通常保留了供體抗體的預期性質，包括但不限於，抗原特異性、親和性、反應性等。供體抗體可以是有預期性質（例如，抗原特異性、親和性、反應性等）的小鼠、大鼠、兔或非人靈長類動物（例如，食蟹猴）抗體。As used herein, the term "humanized antibody" refers to a genetically engineered non-human antibody whose amino acid sequence has been modified to increase homology with the sequence of a human antibody. Generally speaking, all or part of the CDR region of a humanized antibody is derived from a non-human antibody (donor antibody), and all or part of the non-CDR region (for example, variable region FR and/or constant region) is derived from human source. Immunoglobulin (receptor antibody). Humanized antibodies generally retain the expected properties of the donor antibody, including, but not limited to, antigen specificity, affinity, reactivity, etc. The donor antibody may be a mouse, rat, rabbit, or non-human primate (for example, cynomolgus monkey) antibody with desired properties (for example, antigen specificity, affinity, reactivity, etc.).

在本申請中，本發明抗體的預期性質包括：(1) 特異性識別/結合CLDN18.2（特別是人CLDN18.2）；(2) 介導CLDN18.2內化；(3) 藉由抗體依賴性細胞介導的細胞毒性（ADCC）來誘導殺傷表達人CLDN18.2的細胞；(4) 藉由補體依賴的細胞毒性（CDC）來誘導殺傷表達人CLDN18.2的細胞；(5) 預防和/治療腫瘤的能力。本發明的抗體具有上述預期性質中的一項或多項。In this application, the expected properties of the antibody of the present invention include: (1) specific recognition/binding to CLDN18.2 (especially human CLDN18.2); (2) mediation of CLDN18.2 internalization; (3) by the antibody Dependent cell-mediated cytotoxicity (ADCC) is used to induce and kill cells expressing human CLDN 18.2; (4) Complement-dependent cytotoxicity (CDC) is used to induce and kill cells expressing human CLDN 18.2; (5) Prevention And/the ability to treat tumors. The antibody of the present invention has one or more of the aforementioned expected properties.

本發明的嵌合抗體或人源化抗體可以根據上述製備的鼠單殖株抗體的序列進行製備。編碼重鏈和輕鏈的DNA可以從目標鼠雜交瘤中獲得，並且使用標準分子生物學技術進行工程改造以包含非鼠（例如人）免疫球蛋白序列。The chimeric antibody or humanized antibody of the present invention can be prepared based on the sequence of the murine monoclonal antibody prepared above. DNA encoding the heavy and light chains can be obtained from the target murine hybridoma and engineered using standard molecular biology techniques to contain non-mouse (eg, human) immunoglobulin sequences.

為製備嵌合抗體，可使用本領域已知的方法將鼠免疫球蛋白可變區連接至人免疫球蛋白恆定區（參見例如Cabilly等人的美國專利No.4,816,567）。例如，將編碼VH的DNA可操作的連接至編碼重鏈恆定區的另一DNA分子以獲得全長重鏈基因。人重鏈恆定區基因的序列是本領域已知的（參見例如Kabat, E.A.等人(1991) Sequences of Proteins of Immunological Interest, Fifth Edition, U.S. Department of Health and Human Services, NIH Publication No.91-3242），包含這些區的DNA片段可以藉由標準PCR擴增獲得。重鏈恆定區可以是IgG1、IgG2、IgG3、IgG4、IgA、IgE、IgM或IgD恆定區，但是通常較佳為IgG1或IgG4恆定區。例如，將編碼VL的DNA可操作的連接至編碼輕鏈恆定區CL的另一DNA分子以獲得全長輕鏈基因（以及Fab輕鏈基因）。人輕鏈恆定區基因的序列是本領域已知的（參見例如Kabat，E.A. 等人(1991) Sequences of Proteins of Immunological Interest，Fifth Edition，U.S.Department of Health and Human Services，NIH Publication No.91-3242），包含這些區的DNA片段可以藉由標準PCR擴增獲得。輕鏈恆定區可以是κ或λ恆定區，但通常較佳為κ恆定區。To prepare chimeric antibodies, the murine immunoglobulin variable region can be linked to the human immunoglobulin constant region using methods known in the art (see, for example, U.S. Patent No. 4,816,567 to Cabilly et al.). For example, the DNA encoding VH is operably linked to another DNA molecule encoding the heavy chain constant region to obtain a full-length heavy chain gene. The sequence of the human heavy chain constant region gene is known in the art (see, for example, Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, US Department of Health and Human Services, NIH Publication No. 91-3242 ), DNA fragments containing these regions can be obtained by standard PCR amplification. The heavy chain constant region may be an IgG1, IgG2, IgG3, IgG4, IgA, IgE, IgM, or IgD constant region, but is generally preferably an IgG1 or IgG4 constant region. For example, the DNA encoding VL is operably linked to another DNA molecule encoding the light chain constant region CL to obtain the full-length light chain gene (and the Fab light chain gene). The sequence of the human light chain constant region gene is known in the art (see, for example, Kabat, EA et al. (1991) Sequences of Proteins of Immunological Interest, Fifth Edition, USDepartment of Health and Human Services, NIH Publication No. 91-3242 ), DNA fragments containing these regions can be obtained by standard PCR amplification. The light chain constant region may be a kappa or lambda constant region, but it is usually a kappa constant region.

為製備人源化抗體，可以使用本領域已知的方法將鼠CDR區***人源框架序列（參見Winter的美國專利No.5,225,539；Queen等人的美國專利Nos.5,530,101; 5,585,089; 5,693,762和6,180,370；以及Lo, Benny, K.C., editor, in Antibody Engineering: Methods and Protocols, volume 248, Humana Press, New Jersey, 2004）。或者，還可以利用轉基因動物，其能夠在免疫後不產生內源性免疫球蛋白、並且能夠產生完整人抗體庫。例如，已有報導在嵌合和種系突變小鼠中抗體重鏈連接區(JH)基因的純合缺失可以完全抑制了內源性抗體產生，然後將人種系免疫球蛋白基因陣列轉移到所述種系突變小鼠中將導致該小鼠在遇到抗原刺激時產生人抗體（參見例如，Jakobovits等，1993，Proc.Natl.Acad.Sci.USA 90 ：2551 ；Jakobovits 等，1993，Nature362 ：255-258 ；Bruggermann等，1993，Year in Immunology 7 ：33 ；和Duchosal等，1992，Nature 355 ：258）。上述轉基因動物的非限制性實例包括，HuMAb小鼠（Medarex,Inc.），其含有編碼未重排的人重鏈（μ和γ）和κ輕鏈免疫球蛋白序列的人免疫球蛋白基因微型基因座（miniloci），加之使內源μ和κ鏈基因座失活的靶向突變（參見例如Lonberg等人(1994) Nature 368(6474):856-859）；或攜帶人重鏈轉基因和人輕鏈轉染色體的“KM小鼠^TM ”（參見專利申請WO02/43478）。其他抗體人源化改造的方法還包括噬菌體展示技術（Hoogenboom 等，1991，J.Mol.Biol.227 ：381 ；Marks 等，J.Mol.Biol.1991，222 ：581-597 ；Vaughan 等，1996，Nature Biotech 14 ：309）。To prepare humanized antibodies, mouse CDR regions can be inserted into human framework sequences using methods known in the art (see US Patent No. 5,225,539 to Winter; US Patent Nos. 5,530,101 to Queen et al.; 5,585,089; 5,693,762 and 6,180,370; And Lo, Benny, KC, editor, in Antibody Engineering: Methods and Protocols, volume 248, Humana Press, New Jersey, 2004). Alternatively, transgenic animals can also be used, which can produce no endogenous immunoglobulin after immunization, and can produce a complete human antibody library. For example, it has been reported that the homozygous deletion of the antibody heavy chain joining region (JH) gene in chimeric and germline mutant mice can completely inhibit the production of endogenous antibodies, and then transfer the human germline immunoglobulin gene array to The germline mutant mice will cause the mice to produce human antibodies when they encounter antigenic stimulation (see, for example, Jakobovits et al., 1993, Proc. Natl. Acad. Sci. USA 90: 2551; Jakobovits et al., 1993, Nature 362 :255-258; Bruggermann et al., 1993, Year in Immunology 7:33; and Duchosal et al., 1992, Nature 355:258). Non-limiting examples of the above-mentioned transgenic animals include HuMAb mice (Medarex, Inc.), which contain human immunoglobulin genes encoding unrearranged human heavy chain (μ and γ) and κ light chain immunoglobulin sequences. Locus (miniloci), plus targeted mutations that inactivate the endogenous mu and κ chain loci (see, for example, Lonberg et al. (1994) Nature 368(6474):856-859); or carrying human heavy chain transgenes and human The light chain transchromosome "KM ^MouseTM " (see patent application WO02/43478). Other methods of antibody humanization include phage display technology (Hoogenboom et al., 1991, J. Mol. Biol. 227: 381; Marks et al., J. Mol. Biol. 1991, 222: 581-597; Vaughan et al., 1996 , Nature Biotech 14:309).

如本文中所使用的，術語“胚系抗體基因（germline antibody gene）”或“胚系抗體基因片段（germline antibody gene segment）”是指，存在於生物體的基因組中的編碼免疫球蛋白的序列，其沒有經歷過能夠導致表達特異性免疫球蛋白的遺傳學重排及突變的成熟過程。在本發明中，表述“重鏈胚系基因”是指，編碼免疫球蛋白重鏈的胚系抗體基因或基因片段，其包括V基因(variable)、D基因(diversity)、J基因(joining)和C基因(constant)；類似地，表述“輕鏈胚系基因”是指，編碼免疫球蛋白輕鏈的胚系抗體基因或基因片段，其包括V基因(variable)、J基因(joining)和C基因(constant)。在本發明中，由該胚系抗體基因或胚系抗體基因片段所編碼的氨基酸序列也稱為“胚系序列（germline sequence）”。胚系抗體基因或胚系抗體基因片段及其相應的胚系序列是本領域技術人員熟知的，並且可從專業資料庫（例如，IMGT、UNSWIg、NCBI或VBASE2）獲得或查詢。As used herein, the term "germline antibody gene" or "germline antibody gene segment" refers to a sequence encoding immunoglobulin that exists in the genome of an organism , It has not experienced the maturation process that can lead to genetic rearrangement and mutation of expressing specific immunoglobulins. In the present invention, the expression "heavy chain germline gene" refers to the germline antibody gene or gene fragment encoding the heavy chain of immunoglobulin, which includes V gene (variable), D gene (diversity), and J gene (joining) And C gene (constant); similarly, the expression "light chain germline gene" refers to germline antibody genes or gene fragments encoding immunoglobulin light chains, including V gene (variable), J gene (joining) and C gene (constant). In the present invention, the amino acid sequence encoded by the germline antibody gene or germline antibody gene fragment is also called "germline sequence (germline sequence)". Germline antibody genes or germline antibody gene fragments and their corresponding germline sequences are well known to those skilled in the art and can be obtained or inquired from professional databases (for example, IMGT, UNSWIg, NCBI or VBASE2).

如本文中所使用的，術語“特異性結合”是指，兩分子間的非隨機的結合反應，如抗體和其所針對的抗原之間的反應。特異性結合相互作用的強度或親和力可以該相互作用的平衡解離常數(K_D )表示。在本發明中，術語“K_D ”是指特定抗體-抗原相互作用的解離平衡常數，其用於描述抗體與抗原之間的結合親和力。平衡解離常數越小，抗體-抗原結合越緊密，抗體與抗原之間的親和力越高。在某些實施方式中，特異性結合某抗原的抗體（或對某抗原具有特異性的抗體）是指，抗體以小於約10^-9 M，例如小於約10^-9 M、10^-10 M、10^-11 M或10^-12 M或更小的親和力(K_D )結合該抗原。兩分子間的特異性結合性質可使用本領域公知的方法進行測定，例如使用表面等離子體共振術(SPR)在BIACORE儀中測定。As used herein, the term "specific binding" refers to a non-random binding reaction between two molecules, such as the reaction between an antibody and the antigen against which it is directed. The strength or affinity of a specific binding interaction can be _{expressed by the equilibrium dissociation constant (K D} ) of the interaction. In the present invention, the term "K _D "refers to the dissociation equilibrium constant of a specific antibody-antigen interaction, which is used to describe the binding affinity between the antibody and the antigen. The smaller the equilibrium dissociation constant, the tighter the antibody-antigen binding, and the higher the affinity between the antibody and the antigen. In some embodiments, an antibody that specifically binds to a certain antigen (or an antibody that is specific to a certain antigen) refers to an antibody with a concentration of less than about 10 ^-9 M, for example, less than about 10 ^-9 M, 10 ^-10 M, The affinity (K _D ) of 10 ^-11 M or 10 ^-12 M or less binds to the antigen. The specific binding properties between two molecules can be measured using methods known in the art, for example, using surface plasmon resonance (SPR) in a BIACORE instrument.

如本文中所使用的，術語“細胞毒劑”包括對細胞有害(例如殺死細胞)的任何試劑，例如化療藥物、細菌毒素、植物毒素或放射性同位素等。As used herein, the term "cytotoxic agent" includes any agent that is harmful to cells (e.g., kills cells), such as chemotherapeutic drugs, bacterial toxins, phytotoxins, or radioisotopes.

如本文中所使用的，術語“載體(vector)”是指，可將多聚核苷酸***其中的一種核酸運載工具。當載體能使***的多核苷酸編碼的蛋白獲得表達時，載體稱為表達載體。載體可以藉由轉化，轉導或者轉染導入宿主細胞，使其攜帶的遺傳物質元件在宿主細胞中獲得表達。載體是本領域技術人員公知的，包括但不限於：質粒；噬菌粒；柯斯質粒；人工染色體，例如酵母人工染色體(YAC)、細菌人工染色體(BAC)或P1來源的人工染色體(PAC)；噬菌體如λ噬菌體或M13噬菌體及動物病毒等。可用作載體的動物病毒包括但不限於，逆轉錄酶病毒(包括慢病毒)、腺病毒、腺相關病毒、皰疹病毒(如單純皰疹病毒)、痘病毒、杆狀病毒、乳頭瘤病毒、乳頭多瘤空泡病毒(如SV40)。一種載體可以含有多種控制表達的元件，包括但不限於，啟動子序列、轉錄起始序列、增強子序列、選擇元件及報告基因。另外，載體還可含有複製起始位點。As used herein, the term "vector" refers to a nucleic acid delivery vehicle into which polynucleotides can be inserted. When the vector can express the protein encoded by the inserted polynucleotide, the vector is called an expression vector. The vector can be introduced into the host cell by transformation, transduction or transfection, so that the genetic material elements it carries can be expressed in the host cell. Vectors are well known to those skilled in the art, including but not limited to: plasmids; phagemids; cosmids; artificial chromosomes, such as yeast artificial chromosomes (YAC), bacterial artificial chromosomes (BAC) or P1 derived artificial chromosomes (PAC) ; Phages such as lambda phage or M13 phage and animal viruses. Animal viruses that can be used as vectors include, but are not limited to, retrovirus (including lentivirus), adenovirus, adeno-associated virus, herpes virus (such as herpes simplex virus), poxvirus, baculovirus, papilloma virus , Papillary polyoma vacuole virus (such as SV40). A vector can contain a variety of elements that control expression, including but not limited to promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. In addition, the vector may also contain an origin of replication site.

如本文中所使用的，術語“宿主細胞”是指，可用於導入載體的細胞，其包括但不限於，如大腸桿菌或枯草菌等的原核細胞，如酵母細胞或曲黴菌等的真菌細胞，如S2果蠅細胞或Sf9等的昆蟲細胞，或者如纖維原細胞，CHO細胞，COS細胞，NSO細胞，HeLa細胞，BHK細胞，HEK 293細胞或人細胞等的動物細胞。As used herein, the term "host cell" refers to a cell that can be used to introduce a vector, which includes, but is not limited to, prokaryotic cells such as Escherichia coli or subtilis, fungal cells such as yeast cells or Aspergillus, etc. Insect cells such as S2 Drosophila cells or Sf9, or animal cells such as fibroblasts, CHO cells, COS cells, NSO cells, HeLa cells, BHK cells, HEK 293 cells or human cells.

如本文中所使用的，術語“同一性”用於指兩個多肽之間或兩個核酸之間序列的匹配情況。當兩個進行比較的序列中的某個位置都被相同的鹼基或氨基酸單體亞單元佔據時(例如，兩個DNA分子的每一個中的某個位置都被腺嘌呤佔據，或兩個多肽的每一個中的某個位置都被賴氨酸佔據)，那麼各分子在該位置上是同一的。兩個序列之間的“百分數同一性”是由這兩個序列共有的匹配位置數目除以進行比較的位置數目×100的函數。例如，如果兩個序列的10個位置中有6個匹配，那麼這兩個序列具有60%的同一性。例如，DNA序列CTGACT和CAGGTT共有50%的同一性(總共6個位置中有3個位置匹配)。通常，在將兩個序列比對以產生最大同一性時進行比較。這樣的比對可藉由使用，例如，可藉由電腦程式例如Align程式(DNAstar, Inc.)方便地進行的Needleman等人(1970)J. Mol. Biol. 48：443-453的方法來實現。還可使用已整合入ALIGN程式(版本2.0)的E. Meyers和W. Miller (Comput. Appl Biosci.，4:11-17 (1988))的演算法，使用PAM120權重殘基表(weight residue table)、12的缺口長度罰分和4的缺口罰分來測定兩個氨基酸序列之間的百分數同一性。此外，可使用已整合入GCG套裝軟體(可在www.gcg.com上獲得)的GAP程式中的Needleman和Wunsch (J MoI Biol. 48:444-453 (1970))演算法，使用Blossum 62矩陣或PAM250矩陣以及16、14、12、10、8、6或4的缺口權重(gap weight)和1、2、3、4、5或6的長度權重來測定兩個氨基酸序列之間的百分數同一性。As used herein, the term "identity" is used to refer to the matching of sequences between two polypeptides or between two nucleic acids. When a certain position in the two sequences to be compared is occupied by the same base or amino acid monomer subunit (for example, a certain position in each of the two DNA molecules is occupied by adenine, or two A certain position in each of the polypeptides is occupied by lysine), then each molecule is the same at that position. The "percent identity" between two sequences is a function of the number of matching positions shared by the two sequences divided by the number of positions to be compared × 100. For example, if 6 out of 10 positions in two sequences match, then the two sequences have 60% identity. For example, the DNA sequences CTGACT and CAGGTT share 50% identity (3 out of 6 positions match). Generally, the comparison is made when two sequences are aligned to produce maximum identity. Such comparison can be achieved by using, for example, the method of Needleman et al. (1970) J. Mol. Biol. 48:443-453, which is conveniently performed by a computer program such as the Align program (DNAstar, Inc.) . It is also possible to use the algorithm of E. Meyers and W. Miller (Comput. Appl Biosci., 4:11-17 (1988)) that has been integrated into the ALIGN program (version 2.0), using the PAM120 weight residue table (weight residue table). ), a gap length penalty of 12 and a gap penalty of 4 to determine the percent identity between two amino acid sequences. In addition, the Needleman and Wunsch (J MoI Biol. 48:444-453 (1970)) algorithms in the GAP program integrated into the GCG package software (available on www.gcg.com) can be used, and the Blossum 62 matrix can be used Or PAM250 matrix and gap weights of 16, 14, 12, 10, 8, 6 or 4 and length weights of 1, 2, 3, 4, 5 or 6 to determine the percent identity between two amino acid sequences sex.

如本文中所使用的，術語“保守置換”意指不會不利地影響或改變包含氨基酸序列的蛋白/多肽的預期性質的氨基酸置換。例如，可藉由本領域內已知的標準技術例如定點誘變和PCR介導的誘變引入保守置換。保守氨基酸置換包括用具有相似側鏈的氨基酸殘基替代氨基酸殘基的置換，例如用在物理學上或功能上與相應的氨基酸殘基相似(例如具有相似大小、形狀、電荷、化學性質，包括形成共價鍵或氫鍵的能力等)的殘基進行的置換。已在本領域內定義了具有相似側鏈的氨基酸殘基的家族。這些家族包括具有鹼性側鏈(例如，賴氨酸、精氨酸和組氨酸)、酸性側鏈(例如天冬氨酸、谷氨酸)、不帶電荷的極性側鏈(例如甘氨酸、天冬醯胺、穀氨醯胺、絲氨酸、蘇氨酸、酪氨酸、半胱氨酸、色氨酸)、非極性側鏈(例如丙氨酸、纈氨酸、亮氨酸、異亮氨酸、脯氨酸、苯丙氨酸、甲硫氨酸)、β分支側鏈(例如，蘇氨酸、纈氨酸、異亮氨酸)和芳香族側鏈(例如，酪氨酸、苯丙氨酸、色氨酸、組氨酸)的氨基酸。因此，較佳用來自相同側鏈家族的另一個氨基酸殘基替代相應的氨基酸殘基。鑒定氨基酸保守置換的方法在本領域內是熟知的(參見，例如，Brummell等人，Biochem. 32:1180-1187 (1993); Kobayashi等人Protein Eng. 12(10):879-884 (1999); 和Burks等人Proc. Natl Acad. Set USA 94:412-417 (1997)，其藉由引用併入本文)。As used herein, the term "conservative substitution" means an amino acid substitution that does not adversely affect or change the expected properties of the protein/polypeptide comprising the amino acid sequence. For example, conservative substitutions can be introduced by standard techniques known in the art such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions include substitutions of amino acid residues with similar side chains, such as those that are physically or functionally similar to the corresponding amino acid residues (e.g., have similar size, shape, charge, chemical properties, including The ability to form covalent bonds or hydrogen bonds, etc.) is replaced by residues. Families of amino acid residues with similar side chains have been defined in the art. These families include basic side chains (e.g., lysine, arginine, and histidine), acidic side chains (e.g., aspartic acid, glutamate), uncharged polar side chains (e.g., glycine, Asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), non-polar side chains (e.g. alanine, valine, leucine, isoleucine) Acid, proline, phenylalanine, methionine), beta branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, Phenylalanine, tryptophan, histidine) amino acids. Therefore, it is preferable to substitute another amino acid residue from the same side chain family for the corresponding amino acid residue. Methods of identifying conservative substitutions of amino acids are well known in the art (see, for example, Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al. Protein Eng. 12(10):879-884 (1999) ; And Burks et al. Proc. Natl Acad. Set USA 94:412-417 (1997), which is incorporated herein by reference).

本文涉及的二十個常規氨基酸的編寫遵循常規用法。參見例如，Immunology-A Synthesis (2nd Edition, E. S. Golub and D. R. Gren, Eds., Sinauer Associates , Sunderland, Mass. (1991))，其以引用的方式併入本文中。在本發明中，術語“多肽”和“蛋白質”具有相同的含義且可互換使用。並且在本發明中，氨基酸通常用本領域公知的單字母和三字母縮寫來表示。例如，丙氨酸可用A或Ala表示。The preparation of the twenty conventional amino acids involved in this article follows conventional usage. See, for example, Immunology-A Synthesis (2nd Edition, E. S. Golub and D. R. Gren, Eds., Sinauer Associates, Sunderland, Mass. (1991)), which is incorporated herein by reference. In the present invention, the terms "polypeptide" and "protein" have the same meaning and can be used interchangeably. And in the present invention, amino acids are usually represented by one-letter and three-letter abbreviations well known in the art. For example, alanine can be represented by A or Ala.

如本文中所使用的，術語“嵌合抗原受體(CAR)”是指，具有與T細胞受體的一或複數細胞內訊號結構域接合的細胞外抗體衍生的靶向結構域(例如scFv)的工程化的T細胞受體。在本發明中，術語“嵌合抗原受體T細胞”是表達CAR並且具有由該CAR的靶向結構域決定的抗原特異性的T細胞。製造CAR(例如，用於癌症治療)的方法是本領域已知的，可參見例如，Park等人,TrendsBiotechnol.,29:550-557,2011；Grupp等人,NEnglJMed.,368:1509-1518,2013；Han等人,J.HematolOncol.,6:47,2013；PCT專利公開文本WO2012/079000、WO2013/059593；和美國專利公開文本2012/0213783，其全部藉由引用整體併入本文。As used herein, the term "chimeric antigen receptor (CAR)" refers to an extracellular antibody-derived targeting domain (such as scFv ) Of the engineered T cell receptor. In the present invention, the term "chimeric antigen receptor T cell" is a T cell that expresses CAR and has antigen specificity determined by the targeting domain of the CAR. The method of manufacturing CAR (for example, for cancer treatment) is known in the art, see, for example, Park et al., TrendsBiotechnol., 29:550-557, 2011; Grupp et al., NEnglJMed., 368:1509-1518 , 2013; Han et al., J. Hematol Oncol., 6:47, 2013; PCT Patent Publications WO2012/079000, WO2013/059593; and U.S. Patent Publications 2012/0213783, all of which are incorporated herein by reference in their entirety.

如本文中所使用的，術語“藥學上可接受的載體和/或賦形劑”是指在藥理學和/或生理學上與受試者和活性成分相容的載體和/或賦形劑，其是本領域公知的(參見例如Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995)，並且包括但不限於：pH調節劑、表面活性劑、佐劑、離子強度增強劑、稀釋劑、維持滲透壓的試劑、延遲吸收的試劑、防腐劑。例如，pH調節劑包括但不限於磷酸鹽緩衝液。表面活性劑包括但不限於陽離子、陰離子或者非離子型表面活性劑，例如Tween-80。離子強度增強劑包括但不限於氯化鈉。防腐劑包括但不限於各種抗細菌試劑和抗真菌試劑，例如對羥苯甲酸酯、三氯叔丁醇、苯酚、山梨酸等。維持滲透壓的試劑包括但不限於糖、NaCl及其類似物。延遲吸收的試劑包括但不限於單硬脂酸鹽和明膠。稀釋劑包括但不限於水，水性緩衝液(如緩衝鹽水)，醇和多元醇(如甘油)等。防腐劑包括但不限於各種抗細菌試劑和抗真菌試劑，例如硫柳汞、2-苯氧乙醇、對羥苯甲酸酯、三氯叔丁醇、苯酚、山梨酸等。穩定劑具有本領域技術人員通常理解的含義，其能夠穩定藥物中的活性成分的期望活性，包括但不限於谷氨酸鈉、明膠、SPGA、糖類（如山梨醇，甘露醇、澱粉、蔗糖、乳糖、葡聚糖、或葡萄糖）、氨基酸（如谷氨酸、甘氨酸）、蛋白質（如乾燥乳清、白蛋白或酪蛋白）或其降解產物（如乳白蛋白水解物）等。在某些示例性實施方案中，該藥學上可接受的載體或賦形劑包括無菌可注射液體（如水性或非水性懸浮液或溶液）。在某些示例性實施方案中，此類無菌可注射液體選自注射用水(WFI)、抑菌性注射用水(BWFI)、氯化鈉溶液（例如0.9% (w/v)NaCl）、葡萄糖溶液（例如5%葡萄糖）、含有表面活性劑的溶液（例如0.01%聚山梨醇20）、pH緩衝溶液（例如磷酸鹽緩衝溶液）、Ringer氏溶液及其任意組合。As used herein, the term "pharmaceutically acceptable carrier and/or excipient" refers to a carrier and/or excipient that is pharmacologically and/or physiologically compatible with the subject and the active ingredient , Which are well-known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995), and include, but are not limited to: pH adjusters, surfactants, adjuvants, ionic strength Enhancer, diluent, agent to maintain osmotic pressure, agent to delay absorption, preservative. For example, pH adjusting agents include, but are not limited to, phosphate buffer. Surfactants include but are not limited to cationic, anionic or nonionic surfactants, such as Tween-80. Ionic strength enhancers include, but are not limited to, sodium chloride. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid and the like. Agents for maintaining osmotic pressure include, but are not limited to, sugar, NaCl and the like. Agents that delay absorption include, but are not limited to, monostearate and gelatin. Diluents include, but are not limited to, water, aqueous buffers (such as buffered saline), alcohols and polyols (such as glycerol) and the like. Preservatives include, but are not limited to, various antibacterial and antifungal agents, such as thimerosal, 2-phenoxyethanol, parabens, chlorobutanol, phenol, sorbic acid and the like. Stabilizers have the meaning commonly understood by those skilled in the art, which can stabilize the desired activity of the active ingredients in the drug, including but not limited to sodium glutamate, gelatin, SPGA, sugars (such as sorbitol, mannitol, starch, sucrose, Lactose, dextran, or glucose), amino acids (such as glutamic acid, glycine), proteins (such as dried whey, albumin, or casein) or their degradation products (such as lactalbumin hydrolysate), etc. In certain exemplary embodiments, the pharmaceutically acceptable carrier or excipient includes a sterile injectable liquid (such as an aqueous or non-aqueous suspension or solution). In certain exemplary embodiments, such sterile injectable liquid is selected from the group consisting of water for injection (WFI), bacteriostatic water for injection (BWFI), sodium chloride solution (eg 0.9% (w/v) NaCl), glucose solution (Such as 5% glucose), a solution containing a surfactant (such as 0.01% polysorbate 20), a pH buffer solution (such as a phosphate buffer solution), Ringer's solution, and any combination thereof.

如本文中所使用的，術語“預防”是指，為了阻止或延遲疾病或病症或症狀（例如，腫瘤）在受試者體內的發生而實施的方法。如本文中所使用的，術語“治療”是指，為了獲得有益或所需臨床結果而實施的方法。為了本發明的目的，有益或所需的臨床結果包括但不限於，減輕症狀、縮小疾病的範圍、穩定（即，不再惡化）疾病的狀態，延遲或減緩疾病的發展、改善或減輕疾病的狀態、和緩解症狀（無論部分或全部），無論是可檢測或是不可檢測的。此外，“治療”還可以指，與期望的存活期相比（如果未接受治療），延長存活期。As used herein, the term "prevention" refers to a method performed in order to prevent or delay the occurrence of a disease or disorder or symptom (eg, tumor) in a subject. As used herein, the term "treatment" refers to methods performed in order to obtain beneficial or desired clinical results. For the purpose of the present invention, beneficial or desired clinical results include, but are not limited to, alleviating symptoms, narrowing the scope of the disease, stabilizing (ie, no longer worsening) the state of the disease, delaying or slowing the development of the disease, improving or alleviating the disease Status, and relief of symptoms (whether partial or full), whether detectable or undetectable. In addition, "treatment" can also refer to prolonging survival compared to expected survival (if not receiving treatment).

如本文中使用的，術語“受試者”是指哺乳動物，例如靈長類哺乳動物，例如人。在某些實施方式中，該受試者（例如人）患有腫瘤（例如表達CLDN18.2的腫瘤），或者，具有患有上述疾病的風險。As used herein, the term "subject" refers to a mammal, such as a primate mammal, such as a human. In certain embodiments, the subject (e.g., human) has a tumor (e.g., a tumor expressing CLDN 18.2), or is at risk of suffering from the aforementioned diseases.

如本文中所使用的，術語“有效量”是指足以獲得或至少部分獲得期望的效果的量。例如，預防疾病（例如，腫瘤）有效量是指，足以預防，阻止，或延遲疾病（例如，腫瘤）的發生的量；治療疾病有效量是指，足以治癒或至少部分阻止已患有疾病的患者的疾病和其併發症的量。測定這樣的有效量完全在本領域技術人員的能力範圍之內。例如，對於治療用途有效的量將取決於待治療的疾病的嚴重度、患者自己的免疫系統的總體狀態、患者的一般情況例如年齡，體重和性別，藥物的施用方式，以及同時施用的其他治療等等。As used herein, the term "effective amount" refers to an amount sufficient to obtain or at least partially obtain the desired effect. For example, an effective amount for preventing a disease (e.g., tumor) refers to an amount sufficient to prevent, prevent, or delay the occurrence of a disease (e.g., tumor); an effective amount for treating a disease refers to an amount sufficient to cure or at least partially prevent the disease The patient’s disease and the amount of its complications. It is completely within the abilities of those skilled in the art to determine such an effective amount. For example, the effective amount for therapeutic use will depend on the severity of the disease to be treated, the overall state of the patient’s own immune system, the patient’s general conditions such as age, weight and sex, the way the drug is administered, and other treatments that are administered simultaneously etc.

如本文中所使用的，術語“免疫效應細胞”包括具有造血的起源並在免疫應答中起作用的細胞，例如淋巴細胞，例如B細胞和T細胞；天然殺傷細胞；髓樣細胞，例如單核細胞、巨噬細胞、嗜曙紅細胞、肥大細胞、嗜鹼細胞和粒細胞。在某些較佳的實施方案中，該免疫效應細胞是T細胞。As used herein, the term "immune effector cells" includes cells that have hematopoietic origin and play a role in immune responses, such as lymphocytes, such as B cells and T cells; natural killer cells; myeloid cells, such as monocytes Cells, macrophages, eosinophils, mast cells, basophils and granulocytes. In certain preferred embodiments, the immune effector cells are T cells.

如本文中所使用的，術語“轉移”是指，指癌細胞從其原始部位擴散到身體的其他部位。轉移的形成是非常複雜的過程，並依賴於惡性細胞從原發腫瘤中脫離、侵襲胞外基質、透過內皮基底膜以進入體腔和血管以及其後由血液轉運後浸潤靶器官。最後，新腫瘤(即繼發性腫瘤或轉移性腫瘤)在靶部位的生長依賴於血管發生。腫瘤轉移經常甚至在切除原發腫瘤後發生，因為腫瘤細胞或組分可能殘留併發展出轉移潛力。在一個實施方案中，根據本發明的術語“轉移”涉及“遠端轉移”，這涉及遠離原發腫瘤和局部淋巴結系統的轉移。繼發性或轉移性腫瘤的細胞與原始腫瘤中的細胞類似。這意味著例如如果卵巢癌轉移至肝，則該繼發性腫瘤由異常的卵巢細胞(而不是異常的肝細胞)構成。則肝中的腫瘤被稱為轉移性卵巢癌(而不是肝癌)。發明的有益效果As used herein, the term "metastasis" refers to the spread of cancer cells from their original site to other parts of the body. The formation of metastasis is a very complicated process, and it depends on malignant cells to detach from the primary tumor, invade the extracellular matrix, penetrate the endothelial basement membrane to enter the body cavity and blood vessels, and then be transported by blood to infiltrate the target organ. Finally, the growth of new tumors (ie, secondary tumors or metastatic tumors) at the target site depends on angiogenesis. Tumor metastasis often occurs even after removal of the primary tumor, because tumor cells or components may remain and develop metastatic potential. In one embodiment, the term "metastasis" according to the present invention relates to "distal metastasis", which relates to metastasis away from the primary tumor and the local lymph node system. The cells of the secondary or metastatic tumor are similar to the cells of the original tumor. This means that, for example, if ovarian cancer metastasizes to the liver, the secondary tumor is composed of abnormal ovarian cells (rather than abnormal liver cells). Then the tumor in the liver is called metastatic ovarian cancer (rather than liver cancer). The beneficial effects of the invention

與先前技術相比，本發明的技術方案具有以下有益效果：本發明的抗體能夠特異性識別/結合CLDN18.2，並且能夠藉由ADCC和/或CDC來誘導殺傷表達CLDN18.2的細胞（例如，腫瘤細胞）。因此，本發明的抗體具有用於預防和/或治療腫瘤（特別是表達CLDN18.2的腫瘤）的潛力。本發明的人源化抗體不僅保留了親本抗體的功能和性質，而且具有較高的人源化程度，從而可安全地施用給人受試者，而不引發免疫原性反應。尤其令人意外的是，本發明的抗體相比於已知抗CLDN18.2抗體具有明顯提高的親和活性及腫瘤殺傷活性。因此，本發明的抗體（特別是人源化抗體）具有重大的臨床價值。Compared with the prior art, the technical solution of the present invention has the following beneficial effects: The antibody of the present invention can specifically recognize/bind CLDN 18.2, and can induce and kill cells expressing CLDN 18.2 (for example, tumor cells) through ADCC and/or CDC. Therefore, the antibody of the present invention has the potential for preventing and/or treating tumors (especially tumors expressing CLDN 18.2). The humanized antibody of the present invention not only retains the functions and properties of the parent antibody, but also has a higher degree of humanization, so that it can be safely administered to human subjects without triggering an immunogenic reaction. It is particularly surprising that the antibody of the present invention has significantly improved affinity and tumor-killing activity compared with known anti-CLDN18.2 antibodies. Therefore, the antibodies (especially humanized antibodies) of the present invention have great clinical value.

下面將結合附圖和實施例對本發明的實施方案進行詳細描述，但是本領域技術人員將理解，下列附圖和實施例僅用於說明本發明，而不是對本發明的範圍的限定。根據附圖和較佳實施方案的下列詳細描述，本發明的各種目的和有利方面對於本領域技術人員來說將變得可實施。The embodiments of the present invention will be described in detail below with reference to the accompanying drawings and examples. However, those skilled in the art will understand that the following drawings and examples are only used to illustrate the present invention, but not to limit the scope of the present invention. According to the accompanying drawings and the following detailed description of the preferred embodiments, the various objects and advantageous aspects of the present invention will become practicable for those skilled in the art.

序列資訊本發明涉及的部分序列的資訊提供於下面的表1中。表1：序列的描述 SEQ ID NO 描述序列資訊 1 44F7重鏈可變區 QVQLQQSGAELVRPGASVKMSCKASGYTFTSYNMHWVKQAPRQGLEWIGTIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCARGGYYGNSLDYWGQGTTLTVSS 2 44F7輕鏈可變區 DIQMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNAYYYPFTFGSGTKLEIK 3 44F7 HCDR1 GYTFTSY 4 44F7 HCDR2 YPGNGD 5 44F7 HCDR3 GGYYGNSLDY 6 44F7 LCDR1 KSSQSLLNSGNQKNYLT 7 44F7 LCDR2 WASTRES 8 44F7 LCDR3 QNAYYYPFT 9 2F12重鏈可變區 EVQLVESGGGLVKPGGSLKLSCAASGFTFSDYGIHWVRQAPEKGLEWVAYISSGSSTIYYADTVKGRFTISRDNAKNTLFLQMTSLRSEDTAIYYCAKWDRGNCFDYWGQGTTLTVSS 10 2F12輕鏈可變區 DIVLTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKKYLTWYQQKVGQPPKLLIYWASIRECGVPDRFTGSGSGTDFILTISSVQAEDLAVYYCQNAYSYPLTFGAGTKLELK 11 2F12 HCDR1 GFTFSDY 12 2F12 HCDR2 SSGSST 13 2F12 HCDR3 WDRGNCFDY 14 2F12 LCDR1 KSSQSLLNSGNQKKYLT 15 2F12 LCDR2 WASIREC 16 2F12 LCDR3 QNAYSYPLT 17 3F2重鏈可變區 EVMLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWIATINDGGTYTYYPDNVKGRFTISRDNAKNNLYLHMSHLKSDDTAIHYCTRLARGNSMDYWGQGTSVTVSS 18 3F2輕鏈可變區 EVMLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWIATINDGGTYTYYPDNVKGRFTISRDNAKNNLYLHMSHLKSDDTAIHYCTRLARGNSMDYWGQGTSVTVSS 19 3F2 HCDR1 GFTFSTY 20 3F2 HCDR2 NDGGTY 21 3F2 HCDR3 LARGNSMDY 22 3F2 LCDR1 KSSQSLLNSGNQKNYLT 23 3F2 LCDR2 WASTREY 24 3F2 LCDR3 QNNYIYPLT 25 5F9重鏈可變區 QVQLQQSGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGMIHPNSVSTNYNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCAVYFDYWGQGTTLTVSS 26 5F9輕鏈可變區 DIQVTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYASSSLNSGVPKRFSVSRSGSDYSLTISSLESEDFVDYYCLQYATSPPTFGGGTKLEIK 27 5F9 HCDR1 GYTFTSY 28 5F9 HCDR2 HPNSVS 29 5F9 HCDR3 AVYFDY 30 5F9 LCDR1 RASQDIGSSLN 31 5F9 LCDR2 ASSSLNS 32 5F9 LCDR3 LQYATSPPT 33 9F3重鏈可變區 EVRLQQSGPELVKPGASVKIPCKASGYKFTDYNMDWVKQSHGKSLEWIGEINPNNGGTIYNQKFKGKATLTVDKSSSTAYMELRSLTSEDTAVYYCARIYYGNSFAYWGQGTLVTVSS 34 9F3輕鏈可變區 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNLKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYFYPLTFGAGTKLEIK 35 9F3 HCDR1 GYKFTDY 36 9F3 HCDR2 NPNNGG 37 9F3 HCDR3 IYYGNSFAY 38 9F3 LCDR1 KSSQSLLNSGNLKNYLT 39 9F3 LCDR2 WASTRES 40 9F3 LCDR3 QNDYFYPLT 41 10B11重鏈可變區 QVQMKESGAELVKPGASVKISCKASGYAFSTYWMDWVKQRPGKGLEWIGQIYPGNGDTNYNGKFKGKATLTADKSSSTADMQLSSLTSEDSAVYFCARLGYGNSFTYWGQGTLVTVSA 42 10B11輕鏈可變區 DIQVTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGFGTDFTLTISSVQAEDLAVYYCQNAYFYPFTFGSGTKLEIK 43 10B11 HCDR1 GYAFSTY 44 10B11 HCDR2 YPGNGD 45 10B11 HCDR3 LGYGNSFTY 46 10B11 LCDR1 KSSQSLLNSGNQKNYLT 47 10B11 LCDR2 WASTRES 48 10B11 LCDR3 QNAYFYPFT 49 27B5重鏈可變區 QVQLQQPGTELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGNIHPSNGGSNHNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCAPIYYGNSLAYWGHGTLVTVSA 50 27B5輕鏈可變區 DVVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLLYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNSYFYPFTFGSGTKLEIK 51 27B5 HCDR1 GYTFTSY 52 27B5 HCDR2 HPSNGG 53 27B5 HCDR3 IYYGNSLAY 54 27B5 LCDR1 KSSQSLLNSGNQKNYLT 55 27B5 LCDR2 WASTRES 56 27B5 LCDR3 QNSYFYPF 57 37B1重鏈可變區 EVRLQQSGPELVKPGASVKMSCKASGYSFTDYNMHWVKQSHGKSPEWVGYINPNKGGTGYNQKFKGKATLTVNKSSSTANMELRSLTSEDSAVYYCARIWYGNSFAYWGQGTLVTVSA 58 37B1輕鏈可變區 DVVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYFCQNDYFYPFTFGSGTKLEIK 59 37B1 HCDR1 GYSFTDY 60 37B1 HCDR2 NPNKGG 61 37B1 HCDR3 IWYGNSFAY 62 37B1 LCDR1 KSSQSLLNSGNQKNYLT 63 37B1 LCDR2 WASTRES 64 37B1 LCDR3 QNDYFYPFT 65 44A8重鏈可變區 EVRLQQSGPELVKPGASVKMSCKASGYTFTDYNMHWVKQSHGKSLEWIGYINPKNGGIRYNQKFTGKATLTVNKSSSTAYMELRSLTSEDSAVYYCARGGYYGNTLDNWGQGTSVTVSS 66 44A8輕鏈可變區 DIVMTQSPSSLTLTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTSESGVPDRFTGSGSGTDFTLTISSVQAEDLAIYYCQNAYFYPWTFGGGTKLEIK 67 44A8 HCDR1 GYTFTDY 68 44A8 HCDR2 NPKNGG 69 44A8 HCDR3 GGYYGNTLDN 70 44A8 LCDR1 KSSQSLLNSGNQKNYLT 71 44A8 LCDR2 WASTSES 72 44A8 LCDR3 QNAYFYPWT 73 1D10重鏈可變區 QVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGMIYPNSGSINYNEKFKNKATLTVDKSSSTAYMQLSSLTSEDSAVYFCSVYFDYWGQGTTLTVSS 74 1D10輕鏈可變區 DIQGTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYATSSLDSGVPKRFSVSRSGSDYSLTISSLESEDFVDYYCLQYASSPPTFGGGTKLEIK 75 1D10 HCDR1 GYTFTSY 76 1D10 HCDR2 YPNSGS 77 1D10 HCDR3 YFDY 78 1D10 LCDR1 QDIGSSLN 79 1D10 LCDR2 ATSSLDS 80 1D10 LCDR3 LQYASSPPT 81 人IgG1重鏈恆定區 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 82 人κ輕鏈恆定區 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 83 CLDN18.2-ECL1 DNA GACATGTGGAGCACCCAGGACCTGTACGACAACCCCGTGACCAGCGTGTTCCAGTACGAGGGCCTGTGGAGGAGCTGCGTGAGGCAGAGCAGCGGCTTCACCGAGTGCAGGCCCTACTTCACCATCCTGGGCCTGCCCGCCATGCTGCAGGCCGTGAGG 84 CLDN18.2-ECL1-C3d DNA GACCAGTGGAGCACCCAAGACTTGTACAACAACCCCGTAACAGCTGTTTTCAACTACCAGGGGCTGTGGCGCTCCTGTGTCCGAGAGAGCTCTGGCTTCACCGAGTGCCGGGGCTACTTCACCCTGCTGGGGCTGCCAGCCATGCTGCAGGCAGTGCGAGGCAGCGGCAGCGGCGGCGGCGGCAGCGGCGGCGGCGGCAGCGGCAGCCACCTGATCGTGACCCCCGCCGGCTGCGGCGAGCAGAACATGATCGGCATGACCCCCACCGTGATCGCCGTGCACTACCTGGACCAGACCGAGCAGTGGGAGAAGTTCGGCATCGAGAAGAGGCAGGAGGCCCTGGAGCTGATCAAGAAGGGCTACACCCAGCAGCTGGCCTTCAAGCAGCCCAGCAGCGCCTACGCCGCCTTCAACAACAGGCCCCCCAGCACCTGGCTGACCGCCTACGTGGTGAAGGTGTTCAGCCTGGCCGCCAACCTGATCGCCATCGACAGCCACGTGCTGTGCGGCGCCGTGAAGTGGCTGATCCTGGAGAAGCAGAAGCCCGACGGCGTGTTCCAGGAGGACGGCCCCGTGATCCACCAGGAGATGATCGGCGGCTTCAGGAACGCCAAGGAGGCCGACGTGAGCCTGACCGCCTTCGTGCTGATCGCCCTGCAGGAGGCCAGGGACATCTGCGAGGGCCAGGTGAACAGCCTGCCCGGCAGCATCAACAAGGCCGGCGAGTACATCGAGGCCAGCTACATGAACCTGCAGAGGCCCTACACCGTGGCCATCGCCGGCTACGCCCTGGCCCTGATGAACAAGCTGGAGGAGCCCTACCTGGGCAAGTTCCTGAACACCGCCAAGGACAGGAACAGGTGGGAGGAGCCCGACCAGCAGCTGTACAACGTGGAGGCCACCAGCTACGCCCTGCTGGCCCTGCTGCTGCTGAAGGACTTCGACAGCGTGCCCCCCGTGGTGAGGTGGCTGAACGAGCAGAGGTACTACGGCGGCGGCTACGGCAGCACCCAGGCCACCTTCATGGTGTTCCAGGCCCTGGCCCAGTACCAGACCGACGTGCCCGACCACAAGGACCTGAACATGGACGTGAGCTTCCACCTGCCCAGCAGGGGCAGCGAGGAGTTC 85 hCLDN18.2 DNA ATGGCCGTGACCGCCTGCCAGGGCCTGGGCTTCGTGGTGAGCCTGATCGGCATCGCCGGCATCATCGCCGCCACCTGCATGGACCAGTGGAGCACCCAGGACCTGTACAACAACCCCGTGACCGCCGTGTTCAACTACCAGGGCCTGTGGAGGAGCTGCGTGAGGGAGAGCAGCGGCTTCACCGAGTGCAGGGGCTACTTCACCCTGCTGGGCCTGCCCGCCATGCTGCAGGCCGTGAGGGCCCTGATGATCGTGGGCATCGTGCTGGGCGCCATCGGCCTGCTGGTGAGCATCTTCGCCCTGAAGTGCATCAGGATCGGCAGCATGGAGGACAGCGCCAAGGCCAACATGACCCTGACCAGCGGCATCATGTTCATCGTGAGCGGCCTGTGCGCCATCGCCGGCGTGAGCGTGTTCGCCAACATGCTGGTGACCAACTTCTGGATGAGCACCGCCAACATGTACACCGGCATGGGCGGCATGGTGCAGACCGTGCAGACCAGGTACACCTTCGGCGCCGCCCTGTTCGTGGGCTGGGTGGCCGGCGGCCTGACCCTGATCGGCGGCGTGATGATGTGCATCGCCTGCAGGGGCCTGGCCCCCGAGGAGACCAACTACAAGGCCGTGAGCTACCACGCCAGCGGCCACAGCGTGGCCTACAAGCCCGGCGGCTTCAAGGCCAGCACCGGCTTCGGCAGCAACACCAAGAACAAGAAGATCTACGACGGCGGCGCCAGGACCGAGGACGAGGTGCAGAGCTACCCCAGCAAGCACGACTACGTG 86 人源CLDN18.2氨基酸序列 MAVTACQGLGFVVSLIGIAGIIAATCMDQWSTQDLYNNPVTAVFNYQGLWRSCVRESSGFTECRGYFTLLGLPAMLQAVRALMIVGIVLGAIGLLVSIFALKCIRIGSMEDSAKANMTLTSGIMFIVSGLCAIAGVSVFANMLVTNFWMSTANMYTGMGGMVQTVQTRYTFGAALFVGWVAGGLTLIGGVMMCIACRGLAPEETNYKAVSYHASGHSVAYKPGGFKASTGFGSNTKNKKIYDGGARTEDEVQSYPSKHDYV 87 小鼠CLDN18.2氨基酸序列 MSVTACQGLGFVVSLIGFAGIIAATCMDQWSTQDLYNNPVTAVFNYQGLWRSCVRESSGFTECRGYFTLLGLPAMLQAVRALMIVGIVLGVIGILVSIFALKCIRIGSMDDSAKAKMTLTSGILFIISGISAIIGVSVFANMLVTNFWMSTANMYSGMGGMGGMVQTVQTRYTFGAALFVGWVAGGLTLIGGVMMCIACRGLTPDDSNFKAVSYHASGQNVAYRPGGFKASTGFGSNTRNKKIYDGGARTEDDEQSHPTKYDYV 88 人源CLDN 18.1氨基酸序列 MSTTTCQVVAFLLSILGLAGCIAATGMDMWSTQDLYDNPVTSVFQYEGLWRSCVRQSSGFTECRPYFTILGLPAMLQAVRALMIVGIVLGAIGLLVSIFALKCIRIGSMEDSAKANMTLTSGIMFIVSGLCAIAGVSVFANMLVTNFWMSTANMYTGMGGMVQTVQTRYTFGAALFVGWVAGGLTLIGGVMMCIACRGLAPEETNYKAVSYHASGHSVAYKPGGFKASTGFGSNTKNKKIYDGGARTEDEVQSYPSKHDYV 89 175D10重鏈氨基酸序列 QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 90 175D10輕鏈氨基酸序列 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 91 7004-09hu15重鏈可變區 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGTIYPGNGDTSYNQKFQGRVTMTRDKSTSTVYMELSSLRSEDTAVYFCARGGYYGNSLDYWGQGTLVTVS 92 7004-09hu15輕鏈可變區 DIVMTQSPDSLAVSLGERATINCKSSQSVLNSGNQKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNAYYYPFTFGQGTKLEIK 93 7004-09hu15 HCDR1 GYTFTSY 94 7004-09hu15 HCDR2 YPGNGD 95 7004-09hu15 HCDR3 GGYYGNSLDY 96 7004-09hu15 LCDR1 KSSQSVLNSGNQKNYLA 97 7004-09hu15 LCDR2 WASTRES 98 7004-09hu15 LCDR3 QNAYYYPFT 99 7004-09hu09重鏈可變區 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGTIYPGNGDTSYNQKFQGRVTMTRDKSTSTVYMELSSLRSEDTAVYFCARGGYYGNSLDYWGQGTLVTVS 100 7004-09hu09輕鏈可變區 DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNAYYYPFTFGQGTKLEIK 101 7004-09hu10重鏈可變區 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGTIYPGNGDTSYNQKFQGRVTMTRDKSTSTVYMELSSLRSEDTAVYFCARGGYYGNSLDYWGQGTLVTVS 102 7004-09hu10輕鏈可變區 DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNAYYYPFTFGQGTKLEIK 103 1D10重鏈可變區編碼核酸序列 CAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGTTGTCCTGCAAGGCTTCTGGCTACACTTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAATGATTTATCCTAATAGTGGTAGCATTAACTACAATGAGAAGTTCAAGAACAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTTCTGTCTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 104 1D10輕鏈可變區編碼核酸序列 GACATCCAGGGGACACAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTACGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGTCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGACTATTACTGTCTACAATATGCTAGTTCTCCTCCGACGTTCGGTGGAGGCACCAAACTGGAAATCAAA 105 2F12重鏈可變區編碼核酸序列 GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCTGCCTCTGGATTCACTTTCAGTGACTATGGGATCCACTGGGTTCGTCAGGCTCCAGAGAAGGGGCTGGAGTGGGTTGCATACATTAGTAGTGGCAGTAGTACCATCTACTATGCAGACACAGTGAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTTCCTGCAAATGACCAGTCTGAGGTCTGAGGACACGGCCATATATTACTGTGCAAAGTGGGACCGGGGTAACTGCTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 106 2F12輕鏈可變區編碼核酸序列 GACATTGTGTTGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAAATACTTGACCTGGTATCAGCAGAAAGTAGGGCAGCCTCCTAAACTGTTGATTTACTGGGCATCCATTAGGGAATGTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCATTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGCTTATAGTTATCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA 107 3F2重鏈可變區編碼核酸序列 GAGGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGATCGCAACCATTAATGATGGTGGTACTTACACCTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTGTACCTACACATGAGCCATCTGAAGTCTGACGACACAGCCATCCATTACTGTACAAGACTAGCGAGGGGGAATTCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA 108 3F2輕鏈可變區編碼核酸序列 GAGGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGATCGCAACCATTAATGATGGTGGTACTTACACCTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTGTACCTACACATGAGCCATCTGAAGTCTGACGACACAGCCATCCATTACTGTACAAGACTAGCGAGGGGGAATTCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA 109 5F9重鏈可變區編碼核酸序列 CAGGTCCAACTGCAGCAGTCTGGGGCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGTTGTCCTGCAAGGCTTCTGGCTACACTTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAATGATTCATCCTAATAGTGTTAGTACTAACTACAATGAGAAGTTCAAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCTGTCTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 110 5F9輕鏈可變區編碼核酸序列 GACATCCAGGTGACACAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTACGCCTCATCCAGTTTAAATTCTGGTGTCCCCAAAAGGTTCAGTGTCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGACTATTACTGTCTACAATATGCTACTTCTCCTCCGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA 111 9F3重鏈可變區編碼核酸序列 GAGGTTCGGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATACCCTGCAAGGCTTCTGGATACAAATTCACTGACTACAACATGGACTGGGTGAAGCAGAGCCATGGAAAGAGCCTTGAGTGGATTGGAGAAATTAATCCTAACAATGGTGGTACTATCTACAACCAGAAGTTCAAGGGCAAGGCCACATTGACTGTAGACAAGTCCTCCAGCACAGCCTACATGGAGCTCCGCAGCCTGACATCTGAGGACACTGCAGTCTATTACTGTGCAAGAATTTACTATGGTAACTCCTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTTCA 112 9F3輕鏈可變區編碼核酸序列 GACATTGTGATGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCTAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTTTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGATTATTTTTATCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAAATCAAA 113 10B11重鏈可變區編碼核酸序列 CAGGTGCAAATGAAGGAGTCTGGGGCTGAGCTGGTGAAGCCTGGGGCCTCAGTGAAGATTTCCTGCAAAGCTTCTGGCTACGCATTCAGTACCTACTGGATGGACTGGGTGAAGCAGAGGCCTGGAAAGGGTCTTGAGTGGATTGGACAGATTTATCCTGGAAATGGTGATACTAACTACAACGGAAAGTTCAAGGGCAAGGCCACACTGACTGCAGACAAATCCTCCAGCACAGCCGACATGCAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTTCTGTGCAAGATTGGGGTATGGTAACTCGTTTACTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA 114 10B11輕鏈可變區編碼核酸序列 GACATCCAGGTGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATTTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGCTTATTTTTATCCATTCACGTTCGGCTCGGGGACAAAGCTGGAAATCAAA 115 27B5重鏈可變區編碼核酸序列 CAGGTCCAACTGCAGCAGCCTGGGACCGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTCATCCTAGCAATGGTGGTAGTAACCACAATGAGAAGTTCAAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGCGCCTATCTACTATGGTAACTCGCTTGCTTATTGGGGCCACGGGACTCTGGTCACTGTCTCTGCA 116 27B5輕鏈可變區編碼核酸序列 GATGTTGTGATGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTATTGCTCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATAGTTATTTTTATCCATTCACGTTCGGCTCGGGGACAAAGCTGGAAATCAAA 117 37B1重鏈可變區編碼核酸序列 GAGGTTCGGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACTCATTCACTGACTACAACATGCACTGGGTGAAGCAGAGCCATGGAAAGAGCCCTGAGTGGGTTGGATATATTAACCCTAACAAGGGTGGTACTGGCTACAACCAGAAGTTCAAGGGCAAGGCCACATTGACTGTAAACAAGTCCTCCAGCACAGCCAACATGGAGCTCCGCAGCCTGACATCGGAGGATTCCGCAGTCTATTACTGTGCACGGATATGGTATGGTAATTCGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCAGCA 118 37B1輕鏈可變區編碼核酸序列 GATGTTGTGATGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTTCTGTCAGAATGATTATTTTTATCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA 119 44A8重鏈可變區編碼核酸序列 GAGGTTCGGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACATTCACTGACTACAACATGCACTGGGTGAAACAGAGCCATGGAAAGAGCCTTGAGTGGATTGGATATATTAACCCTAAGAATGGTGGTATTAGATACAACCAGAAGTTCACGGGCAAGGCCACATTGACTGTAAACAAGTCCTCCAGCACAGCCTACATGGAGCTCCGCAGCCTGACATCGGAGGATTCTGCAGTCTATTACTGTGCAAGAGGGGGTTACTACGGTAATACTTTGGACAACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA 120 44A8輕鏈可變區編碼核酸序列 GACATTGTGATGACACAGTCTCCATCCTCCCTGACTCTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGTGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAATTTATTACTGTCAGAATGCTTATTTTTATCCGTGGACGTTCGGTGGAGGCACCAAACTGGAGATCAAA 121 44F7重鏈可變區編碼核酸序列 CAGGTCCAACTGCAGCAGTCTGGGGCTGAGCTGGTGAGGCCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACATTTACCAGTTACAATATGCACTGGGTAAAGCAGGCACCTAGACAGGGCCTGGAATGGATTGGAACTATTTATCCAGGAAATGGTGATACTTCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTGCAAGAGGGGGCTACTATGGTAACTCTCTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 122 44F7輕鏈可變區編碼核酸序列 GACATCCAGATGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGCTTATTATTATCCATTCACGTTCGGCTCGGGGACAAAGCTGGAGATCAAA 123 7004-09hu15重鏈可變區編碼核酸序列 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAACTATCTATCCTGGTAATGGTGATACAAGCTACAACCAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGACAAGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTTCTGTGCGAGAGGCGGGTATTATGGGAACAGTCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCC 124 7004-09hu15輕鏈可變區編碼核酸序列 GACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCGGCAACCAGAAGAACTACTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCGTCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATTAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGAATGCATATTACTACCCCTTCACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA 125 7004-09hu09重鏈可變區編碼核酸序列 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACAACATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAACTATCTATCCTGGTAATGGTGATACAAGCTACAACCAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGACAAGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTTCTGTGCGAGAGGCGGGTATTATGGGAACAGTCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCC 126 7004-09hu09輕鏈可變區編碼核酸序列 GACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTCTTTTAAACAGTGGCAACCAGAAGAACTATTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCAGGAACAGACTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGAATGCATATTACTACCCGTTCACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA 127 7004-09hu10重鏈可變區編碼核酸序列 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAACTATCTATCCTGGTAATGGTGATACAAGCTACAACCAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGACAAGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTTCTGTGCGAGAGGCGGGTATTATGGGAACAGTCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCC 128 7004-09hu10輕鏈可變區編碼核酸序列 GACATCGTGATGACCCAGTCTCCAGACTCCCTGGCCGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTCTTTTAAACAGCGGCAACCAGAAGAACTACTTAACTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCGTCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCTCCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAAAATGCATACTACTACCCGTTCACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA Sequence Information The partial sequence information involved in the present invention is provided in Table 1 below. Table 1: Description of the sequence SEQ ID NO describe Sequence Information 1 44F7 heavy chain variable region QVQLQQSGAELVRPGASVKMSCKASGYTFTSYNMHWVKQAPRQGLEWIGTIYPGNGDTSYNQKFKGKATLTVDKSSSTAYMQLSSLTSEDSAVYFCARGGYYGNSLDYWGQGTTLTVSS 2 44F7 light chain variable region DIQMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNAYYYPFTFGSGTKLEIK 3 44F7 HCDR1 GYTFTSY 4 44F7 HCDR2 YPGNGD 5 44F7 HCDR3 GGYYGNSLDY 6 44F7 LCDR1 KSSQSLLNSGNQKNYLT 7 44F7 LCDR2 WASTRES 8 44F7 LCDR3 QNAYYYPFT 9 2F12 heavy chain variable region EVQLVESGGGLVKPGGSLKLSCAASGFTFSDYGIHWVRQAPEKGLEWVAYISSGSSTIYYADTVKGRFTISRDNAKNTLFLQMTSLRSEDTAIYYCAKWDRGNCFDYWGQGTTLTVSS 10 2F12 light chain variable region DIVLTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKKYLTWYQQKVGQPPKLLIYWASIRECGVPDRFTGSGSGTDFILTISSVQAEDLAVYYCQNAYSYPLTFGAGTKLELK 11 2F12 HCDR1 GFTFSDY 12 2F12 HCDR2 SSGSST 13 2F12 HCDR3 WDRGNCFDY 14 2F12 LCDR1 KSSQSLLNSGNQKKYLT 15 2F12 LCDR2 WASIREC 16 2F12 LCDR3 QNAYSYPLT 17 3F2 heavy chain variable region EVMLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWIATINDGGTYTYYPDNVKGRFTISRDNAKNNLYLHMSHLKSDDTAIHYCTRLARGNSMDYWGQGTSVTVSS 18 3F2 light chain variable region EVMLVESGGGLVKPGGSLKLSCAASGFTFSTYAMSWVRQTPEKRLEWIATINDGGTYTYYPDNVKGRFTISRDNAKNNLYLHMSHLKSDDTAIHYCTRLARGNSMDYWGQGTSVTVSS 19 3F2 HCDR1 GFTFSTY 20 3F2 HCDR2 NDGGTY twenty one 3F2 HCDR3 LARGNSMDY twenty two 3F2 LCDR1 KSSQSLLNSGNQKNYLT twenty three 3F2 LCDR2 WASTREY twenty four 3F2 LCDR3 QNNYIYPLT 25 5F9 heavy chain variable region QVQLQQSGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGMIHPNSVSTNYNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCAVYFDYWGQGTTLTVSS 26 5F9 light chain variable region DIQVTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYASSSLNSGVPKRFSVSRSGSDYSLTISSLESEDFVDYYCLQYATSPPTFGGGTKLEIK 27 5F9 HCDR1 GYTFTSY 28 5F9 HCDR2 HPNSVS 29 5F9 HCDR3 AVYFDY 30 5F9 LCDR1 RASQDIGSSLN 31 5F9 LCDR2 ASSSLNS 32 5F9 LCDR3 LQYATSPPT 33 9F3 heavy chain variable region EVRLQQSGPELVKPGASVKIPCKASGYKFTDYNMDWVKQSHGKSLEWIGEINPNNGGTIYNQKFKGKATLTVDKSSSTAYMELRSLTSEDTAVYYCARIYYGNSFAYWGQGTLVTVSS 34 9F3 light chain variable region DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNLKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYFYPLTFGAGTKLEIK 35 9F3 HCDR1 GYKFTDY 36 9F3 HCDR2 NPNNGG 37 9F3 HCDR3 IYYGNSFAY 38 9F3 LCDR1 KSSQSLLNSGNLKNYLT 39 9F3 LCDR2 WASTRES 40 9F3 LCDR3 QNDYFYPLT 41 10B11 heavy chain variable region QVQMKESGAELVKPGASVKISCKASGYAFSTYWMDWVKQRPGKGLEWIGQIYPGNGDTNYNGKFKGKATLTADKSSSTADMQLSSLTSEDSAVYFCARLGYGNSFTYWGQGTLVTVSA 42 10B11 light chain variable region DIQVTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGFGTDFTLTISSVQAEDLAVYYCQNAYFYPFTFGSGTKLEIK 43 10B11 HCDR1 GYAFSTY 44 10B11 HCDR2 YPGNGD 45 10B11 HCDR3 LGYGNSFTY 46 10B11 LCDR1 KSSQSLLNSGNQKNYLT 47 10B11 LCDR2 WASTRES 48 10B11 LCDR3 QNAYFYPFT 49 27B5 heavy chain variable region QVQLQQPGTELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGNIHPSNGGSNHNEKFKSKATLTVDKSSSTAYMQLSSLTSEDSAVYYCAPIYYGNSLAYWGHGTLVTVSA 50 27B5 light chain variable region DVVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLLYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNSYFYPFTFGSGTKLEIK 51 27B5 HCDR1 GYTFTSY 52 27B5 HCDR2 HPSNGG 53 27B5 HCDR3 IYYGNSLAY 54 27B5 LCDR1 KSSQSLLNSGNQKNYLT 55 27B5 LCDR2 WASTRES 56 27B5 LCDR3 QNSYFYPF 57 37B1 heavy chain variable region EVRLQQSGPELVKPGASVKMSCKASGYSFTDYNMHWVKQSHGKSPEWVGYINPNKGGTGYNQKFKGKATLTVNKSSSTANMELRSLTSEDSAVYYCARIWYGNSFAYWGQGTLVTVSA 58 37B1 light chain variable region DVVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYFCQNDYFYPFTFGSGTKLEIK 59 37B1 HCDR1 GYSFTDY 60 37B1 HCDR2 NPNKGG 61 37B1 HCDR3 IWYGNSFAY 62 37B1 LCDR1 KSSQSLLNSGNQKNYLT 63 37B1 LCDR2 WASTRES 64 37B1 LCDR3 QNDYFYPFT 65 44A8 heavy chain variable region EVRLQQSGPELVKPGASVKMSCKASGYTFTDYNMHWVKQSHGKSLEWIGYINPKNGGIRYNQKFTGKATLTVNKSSSTAYMELRSLTSEDSAVYYCARGGYYGNTLDNWGQGTSVTVSS 66 44A8 light chain variable region DIVMTQSPSSLTLTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTSESGVPDRFTGSGSGTDFTLTISSVQAEDLAIYYCQNAYFYPWTFGGGTKLEIK 67 44A8 HCDR1 GYTFTDY 68 44A8 HCDR2 NPKNGG 69 44A8 HCDR3 GGYYGNTLDN 70 44A8 LCDR1 KSSQSLLNSGNQKNYLT 71 44A8 LCDR2 WASTSES 72 44A8 LCDR3 QNAYFYPWT 73 1D10 heavy chain variable region QVQLQQPGAELVKPGASVKLSCKASGYTFTSYWMHWVKQRPGQGLEWIGMIYPNSGSINYNEKFKNKATLTVDKSSSTAYMQLSSLTSEDSAVYFCSVYFDYWGQGTTLTVSS 74 1D10 light chain variable region DIQGTQSPSSLSASLGERVSLTCRASQDIGSSLNWLQQEPDGTIKRLIYATSSLDSGVPKRFSVSRSGSDYSLTISSLESEDFVDYYCLQYASSPPTFGGGTKLEIK 75 1D10 HCDR1 GYTFTSY 76 1D10 HCDR2 YPNSGS 77 1D10 HCDR3 YFDY 78 1D10 LCDR1 QDIGSSLN 79 1D10 LCDR2 ATSSLDS 80 1D10 LCDR3 LQYASSPPT 81 Human IgG1 heavy chain constant region ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 82 Human kappa light chain constant region RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 83 CLDN18.2-ECL1 DNA GACATGTGGAGCACCCAGGACCTGTACGACAACCCCGTGACCAGCGTGTTCCAGTACGAGGGCCTGTGGAGGAGCTGCGTGAGGCAGAGCAGCGGCTTCACCGAGTGCAGGCCCTACTTCACCATCCTGGGCCTGCCCGCCATGCTGCAGGCCGTGAGG 84 CLDN18.2-ECL1-C3d DNA 85 hCLDN18.2 DNA 86 Human CLDN18.2 amino acid sequence MAVTACQGLGFVVSLIGIAGIIAATCMDQWSTQDLYNNPVTAVFNYQGLWRSCVRESSGFTECRGYFTLLGLPAMLQAVRALMIVGIVLGAIGLLVSIFALKCIRIGSMEDSAKANMTLTSGIMFIVSGLCAIAGVSVFANMLVTNFWMSTANMYTGMGGMPGSYGARTSGGAALFVKAGGMVCIAGVAG 87 Mouse CLDN18.2 amino acid sequence MSVTACQGLGFVVSLIGFAGIIAATCMDQWSTQDLYNNPVTAVFNYQGLWRSCVRESSGFTECRGYFTLLGLPAMLQAVRALMIVGIVLGVIGILVSIFALKCIRIGSMDDSAKAKMTLTSGILFIISGISAIIGVSVFANMLVTNFWMSTANMYSGMGGMGGLTYGNAFQVGNAFQVGNFKG 88 Human CLDN 18.1 amino acid sequence MSTTTCQVVAFLLSILGLAGCIAATGMDMWSTQDLYDNPVTSVFQYEGLWRSCVRQSSGFTECRPYFTILGLPAMLQAVRALMIVGIVLGAIGLLVSIFALKCIRIGSMEDSAKANMTLTSGIMFIVSGLCAIAGVSVFANMLVTNFWMSTANMYTGMGGMVQTVQTRYTFGAALFVGSAGGGVHSGVHSGVAGLAG 89 175D10 heavy chain amino acid sequence QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 90 175D10 light chain amino acid sequence DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLSSEKGTASVVCLLNNFYPREATEKVQNSKDSHQSLGSLACESFGSLGSLVSSGSLVSSGSLVSAPVVTSVQDSGSLVSAVSAV 91 7004-09hu15 heavy chain variable region QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGTIYPGNGDTSYNQKFQGRVTMTRDKSTSTVYMELSSLRSEDTAVYFCARGGYYGNSLDYWGQGTLVTVS 92 7004-09hu15 light chain variable region DIVMTQSPDSLAVSLGERATINCKSSQSVLNSGNQKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNAYYYPFTFGQGTKLEIK 93 7004-09hu15 HCDR1 GYTFTSY 94 7004-09hu15 HCDR2 YPGNGD 95 7004-09hu15 HCDR3 GGYYGNSLDY 96 7004-09hu15 LCDR1 KSSQSVLNSGNQKNYLA 97 7004-09hu15 LCDR2 WASTRES 98 7004-09hu15 LCDR3 QNAYYYPFT 99 7004-09hu09 heavy chain variable region QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYNMHWVRQAPGQGLEWMGTIYPGNGDTSYNQKFQGRVTMTRDKSTSTVYMELSSLRSEDTAVYFCARGGYYGNSLDYWGQGTLVTVS 100 7004-09hu09 light chain variable region DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLAWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNAYYYPFTFGQGTKLEIK 101 7004-09hu10 heavy chain variable region QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGTIYPGNGDTSYNQKFQGRVTMTRDKSTSTVYMELSSLRSEDTAVYFCARGGYYGNSLDYWGQGTLVTVS 102 7004-09hu10 light chain variable region DIVMTQSPDSLAVSLGERATINCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNAYYYPFTFGQGTKLEIK 103 1D10 heavy chain variable region encoding nucleic acid sequence CAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGTTGTCCTGCAAGGCTTCTGGCTACACTTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAATGATTTATCCTAATAGTGGTAGCATTAACTACAATGAGAAGTTCAAGAACAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTTCTGTCTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 104 1D10 light chain variable region encoding nucleic acid sequence GACATCCAGGGGACACAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTACGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGTCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGACTATTACTGTCTACAATATGCTAGTTCTCCTCCGACGTTCGGTGGAGGCACCAAACTGGAAATCAAA 105 2F12 heavy chain variable region encoding nucleic acid sequence GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCTGCCTCTGGATTCACTTTCAGTGACTATGGGATCCACTGGGTTCGTCAGGCTCCAGAGAAGGGGCTGGAGTGGGTTGCATACATTAGTAGTGGCAGTAGTACCATCTACTATGCAGACACAGTGAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTTCCTGCAAATGACCAGTCTGAGGTCTGAGGACACGGCCATATATTACTGTGCAAAGTGGGACCGGGGTAACTGCTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 106 2F12 light chain variable region encoding nucleic acid sequence GACATTGTGTTGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAAATACTTGACCTGGTATCAGCAGAAAGTAGGGCAGCCTCCTAAACTGTTGATTTACTGGGCATCCATTAGGGAATGTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCATTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGCTTATAGTTATCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA 107 3F2 heavy chain variable region encoding nucleic acid sequence GAGGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGATCGCAACCATTAATGATGGTGGTACTTACACCTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTGTACCTACACATGAGCCATCTGAAGTCTGACGACACAGCCATCCATTACTGTACAAGACTAGCGAGGGGGAATTCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA 108 3F2 light chain variable region encoding nucleic acid sequence GAGGTGATGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTACCTATGCCATGTCTTGGGTTCGCCAGACTCCGGAAAAGAGGCTGGAGTGGATCGCAACCATTAATGATGGTGGTACTTACACCTACTATCCAGACAATGTAAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACAACCTGTACCTACACATGAGCCATCTGAAGTCTGACGACACAGCCATCCATTACTGTACAAGACTAGCGAGGGGGAATTCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA 109 5F9 heavy chain variable region encoding nucleic acid sequence CAGGTCCAACTGCAGCAGTCTGGGGCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGTTGTCCTGCAAGGCTTCTGGCTACACTTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAATGATTCATCCTAATAGTGTTAGTACTAACTACAATGAGAAGTTCAAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTACTGTGCTGTCTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 110 5F9 light chain variable region coding nucleic acid sequence GACATCCAGGTGACACAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAGTAGCTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTACGCCTCATCCAGTTTAAATTCTGGTGTCCCCAAAAGGTTCAGTGTCAGTAGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGACTATTACTGTCTACAATATGCTACTTCTCCTCCGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA 111 9F3 heavy chain variable region encoding nucleic acid sequence GAGGTTCGGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATACCCTGCAAGGCTTCTGGATACAAATTCACTGACTACAACATGGACTGGGTGAAGCAGAGCCATGGAAAGAGCCTTGAGTGGATTGGAGAAATTAATCCTAACAATGGTGGTACTATCTACAACCAGAAGTTCAAGGGCAAGGCCACATTGACTGTAGACAAGTCCTCCAGCACAGCCTACATGGAGCTCCGCAGCCTGACATCTGAGGACACTGCAGTCTATTACTGTGCAAGAATTTACTATGGTAACTCCTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTTCA 112 9F3 light chain variable region encoding nucleic acid sequence GACATTGTGATGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCTAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTTTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGATTATTTTTATCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAAATCAAA 113 10B11 heavy chain variable region encoding nucleic acid sequence CAGGTGCAAATGAAGGAGTCTGGGGCTGAGCTGGTGAAGCCTGGGGCCTCAGTGAAGATTTCCTGCAAAGCTTCTGGCTACGCATTCAGTACCTACTGGATGGACTGGGTGAAGCAGAGGCCTGGAAAGGGTCTTGAGTGGATTGGACAGATTTATCCTGGAAATGGTGATACTAACTACAACGGAAAGTTCAAGGGCAAGGCCACACTGACTGCAGACAAATCCTCCAGCACAGCCGACATGCAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTTCTGTGCAAGATTGGGGTATGGTAACTCGTTTACTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA 114 10B11 light chain variable region encoding nucleic acid sequence GACATCCAGGTGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATTTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGCTTATTTTTATCCATTCACGTTCGGCTCGGGGACAAAGCTGGAAATCAAA 115 27B5 heavy chain variable region encoding nucleic acid sequence CAGGTCCAACTGCAGCAGCCTGGGACCGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACCAGCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACAAGGCCTTGAGTGGATTGGAAATATTCATCCTAGCAATGGTGGTAGTAACCACAATGAGAAGTTCAAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGCGCCTATCTACTATGGTAACTCGCTTGCTTATTGGGGCCACGGGACTCTGGTCACTGTCTCTGCA 116 27B5 light chain variable region encoding nucleic acid sequence GATGTTGTGATGACCCAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTATTGCTCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATAGTTATTTTTATCCATTCACGTTCGGCTCGGGGACAAAGCTGGAAATCAAA 117 37B1 heavy chain variable region encoding nucleic acid sequence GAGGTTCGGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACTCATTCACTGACTACAACATGCACTGGGTGAAGCAGAGCCATGGAAAGAGCCCTGAGTGGGTTGGATATATTAACCCTAACAAGGGTGGTACTGGCTACAACCAGAAGTTCAAGGGCAAGGCCACATTGACTGTAAACAAGTCCTCCAGCACAGCCAACATGGAGCTCCGCAGCCTGACATCGGAGGATTCCGCAGTCTATTACTGTGCACGGATATGGTATGGTAATTCGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCAGCA 118 37B1 light chain variable region encoding nucleic acid sequence GATGTTGTGATGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTTCTGTCAGAATGATTATTTTTATCCATTCACGTTCGGCTCGGGGACAAAGTTGGAAATAAAA 119 44A8 heavy chain variable region encoding nucleic acid sequence GAGGTTCGGCTGCAACAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACATTCACTGACTACAACATGCACTGGGTGAAACAGAGCCATGGAAAGAGCCTTGAGTGGATTGGATATATTAACCCTAAGAATGGTGGTATTAGATACAACCAGAAGTTCACGGGCAAGGCCACATTGACTGTAAACAAGTCCTCCAGCACAGCCTACATGGAGCTCCGCAGCCTGACATCGGAGGATTCTGCAGTCTATTACTGTGCAAGAGGGGGTTACTACGGTAATACTTTGGACAACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA 120 44A8 light chain variable region encoding nucleic acid sequence GACATTGTGATGACACAGTCTCCATCCTCCCTGACTCTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGGCAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGTGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAATTTATTACTGTCAGAATGCTTATTTTTATCCGTGGACGTTCGGTGGAGGCACCAAACTGGAGATCAAA 121 44F7 heavy chain variable region encoding nucleic acid sequence CAGGTCCAACTGCAGCAGTCTGGGGCTGAGCTGGTGAGGCCTGGGGCCTCAGTGAAGATGTCCTGCAAGGCTTCTGGCTACACATTTACCAGTTACAATATGCACTGGGTAAAGCAGGCACCTAGACAGGGCCTGGAATGGATTGGAACTATTTATCCAGGAAATGGTGATACTTCCTACAATCAGAAGTTCAAGGGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTGCAAGAGGGGGCTACTATGGTAACTCTCTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA 122 44F7 light chain variable region encoding nucleic acid sequence GACATCCAGATGACACAGTCTCCATCCTCCCTGACTGTGACAGCAGGAGAGAAGGTCACTATGAGCTGCAAGTCCAGTCAGAGTCTGTTAAACAGTGGAAATCAAAAGAACTACTTGACCTGGTACCAGCAGAAACCAGGACAGCCTCCTAAACTGTTGATCTACTGGGCATCCACTAGGGAATCTGGGGTCCCTGATCGCTTCACAGGCAGTGGATCTGGAACAGATTTCACTCTCACCATCAGCAGTGTGCAGGCTGAAGACCTGGCAGTTTATTACTGTCAGAATGCTTATTATTATCCATTCACGTTCGGCTCGGGGACAAAGCTGGAGATCAAA 123 7004-09hu15 heavy chain variable region encoding nucleic acid sequence CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAACTATCTATCCTGGTAATGGTGATACAAGCTACAACCAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGACAAGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTTCTGTGCGAGAGGCGGGTATTATGGGAACAGTCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCC 124 7004-09hu15 light chain variable region encoding nucleic acid sequence GACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCGGCAACCAGAAGAACTACTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCGTCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATTAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGAATGCATATTACTACCCCTTCACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA 125 7004-09hu09 heavy chain variable region encoding nucleic acid sequence CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACAACATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAACTATCTATCCTGGTAATGGTGATACAAGCTACAACCAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGACAAGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTTCTGTGCGAGAGGCGGGTATTATGGGAACAGTCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCC 126 7004-09hu09 light chain variable region encoding nucleic acid sequence GACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTCTTTTAAACAGTGGCAACCAGAAGAACTATTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCATCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCAGGAACAGACTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGAATGCATATTACTACCCGTTCACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA 127 7004-09hu10 heavy chain variable region encoding nucleic acid sequence CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGTTTCCTGCAAGGCATCTGGATACACCTTCACCAGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGAACTATCTATCCTGGTAATGGTGATACAAGCTACAACCAGAAGTTCCAGGGCAGAGTCACCATGACCAGGGACAAGTCCACGAGCACAGTCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACGGCCGTGTATTTCTGTGCGAGAGGCGGGTATTATGGGAACAGTCTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCC 128 7004-09hu10 light chain variable region encoding nucleic acid sequence GACATCGTGATGACCCAGTCTCCAGACTCCCTGGCCGTGTCTCTGGGCGAGAGGGCCACCATCAACTGCAAGTCCAGCCAGAGTCTTTTAAACAGCGGCAACCAGAAGAACTACTTAACTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGTTGCTCATTTACTGGGCGTCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCTCCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAAAATGCATACTACTACCCGTTCACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA

現參照下列意在舉例說明本發明(而非限定本發明)的實施例來描述本發明。The invention will now be described with reference to the following examples which are intended to illustrate the invention rather than limit the invention.

除非特別指明，本發明中所使用的分子生物學實驗方法和免疫檢測法，基本上參照J. Sambrook等人，分子殖株：實驗室手冊，第2版，冷泉港實驗室出版社，1989，以及F. M. Ausubel等人，精編分子生物學實驗指南，第3版，John Wiley & Sons, Inc.，1995中所述的方法進行；限制性內切酶的使用依照產品製造商推薦的條件。本領域技術人員知曉，實施例以舉例方式描述本發明，且不意欲限制本發明所要求保護的範圍。實施例1：抗CLDN18.2鼠源抗體的產生Unless otherwise specified, the molecular biology experimental methods and immunoassay methods used in the present invention basically refer to J. Sambrook et al., Molecular Clones: Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press, 1989, And FM Ausubel et al., Compiled Molecular Biology Experiment Guide, 3rd edition, John Wiley & Sons, Inc., 1995; the restriction enzymes are used in accordance with the conditions recommended by the product manufacturer. Those skilled in the art know that the embodiments describe the present invention by way of example, and are not intended to limit the scope of protection claimed by the present invention. Example 1: Production of anti-CLDN18.2 murine antibody

為了獲得抗人CLDN18.2抗體，使用不同的免疫策略（表2）對小鼠（北京維通利華實驗動物技術有限公司，品系代碼216）進行接種，以引發鼠源單抗的產生。其中，抗原包括：表達人CLDN18.2胞外1區編碼核酸序列的表達質粒（CLDN18.2-ECL1 DNA；SEQ ID NO: 83，載體為pcDNA3.1）、表達人CLDN18.2胞外1區-補體C3-編碼核酸序列的表達質粒（CLDN18.2-ECL1-C3d DNA；SEQ ID NO: 84，載體為pcDNA3.1）、表達人CLDN18.2全長編碼核酸序列的表達質粒（hCLDN18.2 DNA；SEQ ID NO: 85，載體為pcDNA3.1）、經轉染表達人源CLDN18.2的中國倉鼠卵巢細胞（CHO-hCLDN18.2）、經轉染表達人源CLDN18.2高表達腎胚細胞（HEK293-hCLDN18.2）。佐劑包括：In vivo-jetPEI（Polyplus Transfection公司，貨號201-50G）、ODN 1826 VacciGrade（InvivoGen公司，貨號vac-1826-1）、完全弗氏佐劑 CFA（InvivoGen公司，貨號vac-cfa-60）。施用途徑包括：肌肉(im)、腹膜內(ip)及皮下(sc)。在加強免疫3天后將免疫小鼠的脾細胞與小鼠骨髓瘤細胞SP2/0使用聚乙二醇法進行融合，得到既能表達抗體又能在體外無限增殖的B細胞融合，並且在HAT選擇培養基中培養。將融合後的雜交瘤細胞鋪在96孔細胞培養板中，並且藉由初級篩選挑選出陽性殖株進行2-3輪次選殖。表2：用於產生單殖株抗體的免疫方案抗原劑量佐劑免疫時長施用途徑 CLDN18.2-ECL1 DNA 100μg 首次，之後每次50μg In vivo-jetPEI 2週一次，共3次 IP CLDN18.2-ECL1 DNA 100μg 首次，之後每次50μg In vivo-jetPEI 3週一次，共3次 IP CHO-hCLDN18.2 5E6 細胞首次CFA，之後 PBS 2週一次，共4次 sc HEK293-hCLDN18.2 5E6 細胞首次CFA，之後 PBS 2週一次，共4次 sc CLDN18.2-ECL1-C3d DNA 100μg每次 In vivo-jetPEI & ODN 1826 2週一次，共4次 IM hCLDN18.2 DNA 100μg每次 In vivo-jetPEI & ODN 1826 2週一次，共4次 IM In order to obtain anti-human CLDN18.2 antibodies, different immunization strategies (Table 2) were used to inoculate mice (Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd., strain code 216) to induce the production of mouse monoclonal antibodies. Among them, the antigen includes: expression plasmid (CLDN18.2-ECL1 DNA; SEQ ID NO: 83, vector pcDNA3.1) expressing the nucleic acid sequence encoding human CLDN18.2 extracellular 1 region, and expressing human CLDN18.2 extracellular 1 region -Complement C3-encoding nucleic acid sequence expression plasmid (CLDN18.2-ECL1-C3d DNA; SEQ ID NO: 84, vector is pcDNA3.1), expression plasmid expressing human CLDN18.2 full-length encoding nucleic acid sequence (hCLDN18.2 DNA ; SEQ ID NO: 85, the vector is pcDNA3.1), Chinese hamster ovary cells transfected to express human CLDN18.2 (CHO-hCLDN18.2), and renal embryo cells transfected to express human CLDN18.2 high expression (HEK293-hCLDN18.2). Adjuvants include: In vivo-jetPEI (Polyplus Transfection company, product number 201-50G), ODN 1826 VacciGrade (InvivoGen company, product number vac-1826-1), complete Freund’s adjuvant CFA (InvivoGen company, product number vac-cfa-60 ). The route of administration includes: intramuscular (im), intraperitoneal (ip) and subcutaneous (sc). Three days after the booster immunization, the spleen cells of the immunized mice were fused with mouse myeloma cells SP2/0 using the polyethylene glycol method to obtain B cells that can express antibodies and proliferate immortally in vitro, and select in HAT Cultivate in medium. The fused hybridoma cells were plated in a 96-well cell culture plate, and positive clones were selected by primary screening for 2-3 rounds of selection. Table 2: Immunization protocols used to produce monoclonal antibodies antigen dose Adjuvant Immunization time Route of administration CLDN18.2-ECL1 DNA 100μg for the first time, then 50μg each time In vivo-jetPEI Once every 2 weeks, 3 times in total IP CLDN18.2-ECL1 DNA 100μg for the first time, then 50μg each time In vivo-jetPEI Once every 3 weeks, 3 times in total IP CHO-hCLDN18.2 5E6 cells First CFA, after PBS Once every 2 weeks, 4 times in total sc HEK293-hCLDN18.2 5E6 cells First CFA, after PBS Once every 2 weeks, 4 times in total sc CLDN18.2-ECL1-C3d DNA 100μg each time In vivo-jetPEI & ODN 1826 Once every 2 weeks, 4 times in total IM hCLDN18.2 DNA 100μg each time In vivo-jetPEI & ODN 1826 Once every 2 weeks, 4 times in total IM

初級篩選：在初級篩選中藉由使用表達人源CLDN18.2的細胞測試生長殖株的上清液對細胞表面CLDN18.2結合能力。藉由用DyLight488山羊抗鼠IgG(Abcam目錄編號ab97015)第二抗體揭示出在上清液中的反應抗體的存在，在全視野細胞掃描分析儀上進行結合能力的評估（參見實施例3）。Primary screening: In the primary screening, cells expressing human-derived CLDN 18.2 are used to test the ability of the supernatant of the growth clone to bind to the cell surface CLDN 18.2. By using the DyLight488 goat anti-mouse IgG (Abcam catalog number ab97015) secondary antibody to reveal the presence of the reactive antibody in the supernatant, the binding ability was evaluated on a full-field cell scanning analyzer (see Example 3).

次級篩選：對上述結合人源CLDN18.2的融合殖株上清液，使用人源CLDN18.表達的細胞測試上清液對細胞表面CLDN18.1結合能力。藉由用DyLight488山羊抗鼠IgG(Abcam目錄編號ab97015)第二抗體揭示出在上清液中的反應抗體的存在，在全視野細胞掃描分析儀上進行結合能力的評估（詳細實驗步驟可參見實施例3）。最終獲得10株陽性雜交瘤單殖株細胞株，然後從其培養上清中分離純化得到以下抗體：1D10、2F12、3F2、5F9、9F3、10B11、27B5、37B1、44A8、44F7。實施例2：抗CLDN18.2鼠源抗體的抗原結合活性評價 2.1 表達CLDN18.2的細胞系的構建Secondary screening: The supernatant of the above-mentioned fusion clone that binds to human CLDN 18.2 is used to test the ability of the supernatant to bind to CLDN 18.1 on the cell surface using cells expressing human CLDN 18. By using the DyLight488 goat anti-mouse IgG (Abcam catalog number ab97015) secondary antibody to reveal the presence of the reactive antibody in the supernatant, the binding ability was evaluated on a full-field cell scanning analyzer (detailed experimental procedures can be found in the implementation Example 3). Finally, 10 positive hybridoma monogenic cell lines were obtained, and the following antibodies were isolated and purified from the culture supernatant: 1D10, 2F12, 3F2, 5F9, 9F3, 10B11, 27B5, 37B1, 44A8, 44F7. Example 2: Evaluation of antigen binding activity of anti-CLDN18.2 murine antibody 2.1 Construction of a cell line expressing CLDN18.2

藉由慢病毒感染和抗性篩選的方法（MOI=3-10，5μg/ml polybrene）在HEK293T細胞（ATCC）、CHOS細胞（Invitrogen）、OCUM-1胃癌腫瘤細胞（南京科佰生物科技有限公司）上分別過表達人源CLDN18.2（SEQ ID NO: 86）或CLDN18.1（SEQ ID NO: 88），或小鼠CLDN18.2（SEQ ID NO: 87）。慢病毒由上海吉凱基因化學技術有限公司提供，細胞感染72小時後加相應抗性繼續培養2-4周，擴增並凍存，以用於後續實驗。 2.2 藉由細胞掃描分析儀檢測鼠源抗體與細胞表面CLDN18.2與CLDN18.1的結合情況By lentivirus infection and resistance screening method (MOI=3-10, 5μg/ml polybrene) in HEK293T cells (ATCC), CHOS cells (Invitrogen), OCUM-1 gastric cancer tumor cells (Nanjing Kebai Biotechnology Co., Ltd.) ) Respectively overexpress human CLDN18.2 (SEQ ID NO: 86) or CLDN18.1 (SEQ ID NO: 88), or mouse CLDN18.2 (SEQ ID NO: 87). The lentivirus was provided by Shanghai Jikai Gene Chemical Technology Co., Ltd. 72 hours after infection, the cells were incubated with corresponding resistance for 2-4 weeks, amplified and frozen for subsequent experiments. 2.2 Detect the binding of mouse-derived antibodies to the cell surface CLDN18.2 and CLDN18.1 by using a cell scanning analyzer

使用重組表達人源CLDN18.2(hCLDN18.2)的HEK293T-hCLDN18.2和OCUM-1-hCLDN18.2，或表達小鼠CLDN18.2（mCLDN18.2）的HEK293T-mCLDN18.2，以及相應的陰性對照細胞系HEK293T；或者使用重組表達人源CLDN18.1(hCLDN18.1)的HEK293T-hCLDN18.1、CHOS-hCLDN18.1，使用DyLight488山羊抗鼠IgG(Abcam目錄編號ab97015)或是DyLight488山羊抗人IgG(Abcam目錄編號ab97003)作為第二抗體，使用以下方法產生滴定結合曲線。Use recombinant HEK293T-hCLDN18.2 and OCUM-1-hCLDN18.2 expressing human CLDN18.2 (hCLDN18.2), or HEK293T-mCLDN18.2 expressing mouse CLDN18.2 (mCLDN18.2), and corresponding Negative control cell line HEK293T; or use recombinant human CLDN18.1 (hCLDN18.1) HEK293T-hCLDN18.1, CHOS-hCLDN18.1, use DyLight488 goat anti-mouse IgG (Abcam catalog number ab97015) or DyLight488 goat anti Human IgG (Abcam catalog number ab97003) was used as the secondary antibody, and the following method was used to generate a titration binding curve.

將10000個細胞平鋪於100μL DMEM+10% FBS/孔中，使用平底96孔板，過夜使細胞貼壁或沉於孔底，第二天去掉上清。藉由將1/3體積(100μL)稀釋於200μL DMEM中來實施3倍梯度稀釋。細胞板的每一孔中加入100μL經稀釋的抗體（篩選時使用融合殖株上清或次選殖上清），相應陰性對照孔加入100μL DMEM，室溫孵育1小時。去掉上清後，每孔加入100μL第二抗體(5μg/mL，稀釋於DMEM中)，室溫孵育0.5小時。染色完成後去掉上清，用PBS清洗一次後每孔加入100μL PBS，然後上機器進行讀數。Spread 10,000 cells in 100μL DMEM+10% FBS/well. Use a flat-bottomed 96-well plate. Allow the cells to adhere to the wall or sink to the bottom of the well overnight. Remove the supernatant the next day. A 3-fold serial dilution was performed by diluting 1/3 volume (100 μL) in 200 μL DMEM. Add 100μL of diluted antibody to each well of the cell plate (use fusion clone supernatant or sub-selection supernatant when screening), add 100μL DMEM to the corresponding negative control well, and incubate for 1 hour at room temperature. After removing the supernatant, add 100 μL of the secondary antibody (5 μg/mL, diluted in DMEM) to each well, and incubate at room temperature for 0.5 hours. After staining, remove the supernatant, wash with PBS once, add 100μL PBS to each well, and then read on the machine.

使用全視野細胞掃描分析儀(Nexcelom公司，型號Celigo® Image Cytometer)對實驗板進行測定讀數。測定時選擇第二抗體對應綠色螢光通道和明場通道同時對孔內細胞進行高速掃描成像，綠色螢光通道得到的成像根據有螢光標記的細胞形態和螢光強度設定參數對對抗體有結合的細胞進行計數，明場通道得到的成像根據細胞形態設定參數對貼壁細胞進行計數，然後兩組資料相除得到和抗體有結合顯示綠色螢光的細胞占貼壁細胞總數的百分比（%螢光細胞），根據該比例判定抗CLDN18.2抗體與表達CLDN18.2細胞的結合活性，百分比越低代表被測定抗CLDN18.2抗體結合細胞表面的CLDN18.2的能力越差，反之百分比越高代表被測定抗CLDN18.2抗體結合細胞表面的CLDN18.2的能力越好。資料分析使用GraphPad。A full-field cell scanning analyzer (Nexcelom company, model Celigo® Image Cytometer) was used to measure and read the test plate. During the measurement, select the green fluorescent channel and bright field channel of the secondary antibody to scan and image the cells in the well at the same time. The imaging obtained by the green fluorescent channel is based on the fluorescently labeled cell morphology and fluorescence intensity setting parameters. The bound cells are counted, and the bright-field channel imaging is based on the cell morphology setting parameters to count the adherent cells, and then the two sets of data are divided to obtain the percentage of cells that bind to the antibody and display green fluorescence to the total number of adherent cells (% Fluorescent cells), based on the ratio to determine the binding activity of the anti-CLDN18.2 antibody to cells expressing CLDN18.2. The lower the percentage, the worse the ability of the tested anti-CLDN18.2 antibody to bind to CLDN18.2 on the cell surface, and vice versa. High means that the ability of the tested anti-CLDN18.2 antibody to bind to CLDN18.2 on the cell surface is better. Data analysis uses GraphPad.

抗CLDN18.2抗體與表達人CLDN18.2的HEK293T、表達鼠CLDN18.2的HEK293T、表達人CLDN18.1的HEK293T和對照HEK293T的結合活性測定結果分別如圖1A-1D所示，橫坐標使用抗體濃度的對數，縱坐標使用和CLDN18.2抗體有結合顯示綠色螢光的細胞占貼壁細胞總數的百分比。進一步根據曲線擬合出抗CLDN18.2抗體結合抗原活性的EC50，其結果如表3所示。其中，參比抗體為175D10（Ganymed Pharmaceuticals AG），揭露於例如CN101312989B，CN103509114B。表3：抗CLDN18.2抗體與CLDN18.2的結合活性測定結果抗體 EC50 (μg/ml) HEK293- hCLDNA18.2 HEK293- mCLDN18.2 OCUM1- hCLDN18.2 HEK293 1D10 0.018 0.691 0.026 N.B. 2F12 0.041 0.036 0.186 N.B. 3F2 0.034 0.059 0.121 N.B. 5F9 0.091 0.179 0.068 N.B. 9F3 0.053 0.043 0.150 N.B. 10B11 0.038 0.035 0.135 N.B. 27B5 0.041 0.033 0.142 N.B. 37B1 0.040 0.032 0.146 N.B. 44A8 0.034 0.026 0.097 N.B. 44F7 0.028 0.026 0.092 N.B. 175D10 1.22 0.81 6.54 N.B. 注：N.B.表示在測定濃度範圍內沒有結合。The binding activity of anti-CLDN18.2 antibody and HEK293T expressing human CLDN18.2, HEK293T expressing murine CLDN18.2, HEK293T expressing human CLDN18.1, and control HEK293T are shown in Figures 1A-1D. The abscissa uses the antibody. The logarithm of the concentration, the ordinate is used and CLDN18.2 antibody binds to the percentage of cells showing green fluorescence to the total number of adherent cells. The EC50 of the antigen binding activity of the anti-CLDN18.2 antibody was further fitted according to the curve, and the results are shown in Table 3. Among them, the reference antibody is 175D10 (Ganymed Pharmaceuticals AG), which is disclosed in, for example, CN101312989B, CN103509114B. Table 3: Results of the determination of the binding activity of anti-CLDN18.2 antibodies to CLDN18.2 antibody EC50 (μg/ml) HEK293- hCLDNA18.2 HEK293- mCLDN18.2 OCUM1- hCLDN18.2 HEK293 1D10 0.018 0.691 0.026 NB 2F12 0.041 0.036 0.186 NB 3F2 0.034 0.059 0.121 NB 5F9 0.091 0.179 0.068 NB 9F3 0.053 0.043 0.150 NB 10B11 0.038 0.035 0.135 NB 27B5 0.041 0.033 0.142 NB 37B1 0.040 0.032 0.146 NB 44A8 0.034 0.026 0.097 NB 44F7 0.028 0.026 0.092 NB 175D10 1.22 0.81 6.54 NB Note: NB means that there is no binding within the measured concentration range.

上述結果顯示，1D10、2F12、3F2、5F9、9F3、10B11、27B5、37B1、44A8、44F7均可結合表達人CLDN18.2的細胞，且顯著優於參比抗體175D10，同時這些抗體不結合不表達CLDN18.2的陰性對照細胞(HEK293T)。The above results show that 1D10, 2F12, 3F2, 5F9, 9F3, 10B11, 27B5, 37B1, 44A8, 44F7 can bind to cells expressing human CLDN18.2, and is significantly better than the reference antibody 175D10, and these antibodies do not bind and do not express CLDN18.2 negative control cells (HEK293T).

抗CLDN18.2抗體與CLDN18.1的結合活性的EC50如表4所示。結果顯示，2F12、3F2、9F3、10B11、27B5、37B1、44A8、44F7均不結合CLDN18.1，顯示出針對CLDN18.2的良好的結合特異性。表4：抗CLDN18.2抗體與CLDN18.1的結合活性測定結果抗體 EC50 μg/ml CHOS- hCLDN18.1 HEK293- hCLDN18.1 1D10 0.092 0.041 2F12 N.B. N.B. 3F2 N.B. 5F9 0.714 9F3 N.B. N.B. 10B11 N.B. N.B. 27B5 N.B. N.B. 37B1 N.B. N.B. 44A8 N.B. N.B. 44F7 N.B. N.B. 175D10 N.B. N.B. 注：N.B.表示在測定濃度範圍內沒有結合，空白表示該抗體未測量此項資料。實施例3：抗CLDN18.2鼠源抗體的序列測定及嵌合抗體的製備 3.1 抗人CLDN18.2鼠源抗體可變區序列的測定The EC50 of the binding activity of the anti-CLDN18.2 antibody to CLDN18.1 is shown in Table 4. The results showed that 2F12, 3F2, 9F3, 10B11, 27B5, 37B1, 44A8, 44F7 did not bind to CLDN 18.1, showing good binding specificity for CLDN 18.2. Table 4: Results of the determination of the binding activity of anti-CLDN18.2 antibodies to CLDN18.1 antibody EC50 μg/ml CHOS- hCLDN18.1 HEK293- hCLDN18.1 1D10 0.092 0.041 2F12 NB NB 3F2 NB 5F9 0.714 9F3 NB NB 10B11 NB NB 27B5 NB NB 37B1 NB NB 44A8 NB NB 44F7 NB NB 175D10 NB NB Note: NB means that there is no binding within the measured concentration range, and a blank means that this antibody has not been measured for this data. Example 3: Sequence determination of anti-CLDN18.2 murine antibody and preparation of chimeric antibody 3.1 Determination of variable region sequence of anti-human CLDN18.2 murine antibody

離心收集雜交瘤細胞，每5-10×10⁶ 個細胞加入1ml TRIzol和0.2ml氯仿，劇烈振盪15秒，室溫放置3分鐘，離心取水相加入0.5ml異丙醇，室溫放置10分鐘後收集沉澱，乙醇洗滌後乾燥得到RNA。在冰浴離心管裡面加入範本RNA和引物，使引物和範本正確配對後進行反轉錄，而後進行PCR擴增。擴增結束後，取4個微量離心管各加入dNTP/ddNTP混合物2.5μl，37℃溫浴5min，備用。在一個空的微量離心管中加入1pmol的PCR擴增產物、10pmol測序引物、2μl 5×測序緩衝液，加雙蒸水至總體積10μl，96℃加熱8min，冰浴泠卻1min，4℃ 10000g離心10s。加入2μl預冷的標記混合物(dCTP、dGTP 、dTTP 各0.75μmol/L)、α-32P-dATP 5μCi、1μl 0.1mol/L DDT、測序酶2U，加水至15μl，混勻後置冰上2min。3.5μl標記反應混合物加入到準備好的4個微量離心管中，37℃溫浴5min。每管各加入4μl終止液。樣品在80℃的水浴中熱變性5min，每一泳道加2μl 加到測序膠上，電泳分離這些片段，收集序列資訊。Collect hybridoma cells by centrifugation, ^{add 1ml TRIzol and 0.2ml chloroform for every 5-10×10 6} cells, shake vigorously for 15 seconds, leave at room temperature for 3 minutes, centrifuge to take the water phase and add 0.5ml isopropanol, leave it at room temperature for 10 minutes The precipitate is collected, washed with ethanol and dried to obtain RNA. Add the template RNA and primers to the ice-bath centrifuge tube, make the primers and the template correctly paired, perform reverse transcription, and then perform PCR amplification. After the amplification, add 2.5μl of dNTP/ddNTP mixture to each of the 4 microcentrifuge tubes, and incubate at 37°C for 5 minutes for use. Add 1pmol PCR amplification product, 10pmol sequencing primer, 2μl 5× sequencing buffer to an empty microcentrifuge tube, add double distilled water to a total volume of 10μl, heat at 96℃ for 8min, cool in ice bath for 1min, 4℃ 10000g Centrifuge for 10 seconds. Add 2μl of pre-chilled labeling mixture (dCTP, dGTP, dTTP each 0.75μmol/L), α-32P-dATP 5μCi, 1μl 0.1mol/L DDT, Sequencing enzyme 2U, add water to 15μl, mix well and place on ice for 2min. Add 3.5 μl of the labeled reaction mixture to the prepared 4 microcentrifuge tubes, and incubate at 37°C for 5 min. Add 4μl stop solution to each tube. The samples were heat-denatured in a water bath at 80°C for 5 minutes, and 2μl per lane was added to the sequencing gel. The fragments were separated by electrophoresis and sequence information was collected.

10株鼠源抗體的VH和VL序列如下表5所示。進一步，還使用Kabat等人描述的方法(Kabat等，Sequences of Proteins of Immunological Interest,第五版，Public Health Service，美國國立衛生研究院，貝塞斯達、馬里蘭州(1991)，第647-669頁)，確定了10株鼠源單抗的CDR序列（表6）。表5: 鼠源抗體輕重鏈可變區氨基酸序列抗體 VH VL SEQ ID NO SEQ ID NO 44F7 1 2 2F12 9 10 3F2 17 18 5F9 25 26 9F3 33 34 10B11 41 42 27B5 49 50 37B1 57 58 44A8 65 66 1D10 73 74 表6: 鼠源抗體CDR的序列抗體 VH (SEQ ID NO:) VL (SEQ ID NO:) CDR1 CDR2 CDR3 CDR1 CDR2 CDR3 44F7 3 4 5 6 7 8 2F12 11 12 13 14 15 16 3F2 19 20 21 22 23 24 5F9 27 28 29 30 31 32 9F3 35 36 37 38 39 40 10B11 43 44 45 46 47 48 27B5 51 52 53 54 55 56 37B1 59 60 61 62 63 64 44A8 67 68 69 70 71 72 1D10 75 76 77 78 79 80 3.2 人-鼠嵌合抗體的製備及抗原結合活性評價The VH and VL sequences of the 10 murine antibodies are shown in Table 5 below. Further, the method described by Kabat et al. (Kabat et al., Sequences of Proteins of Immunological Interest, fifth edition, Public Health Service, National Institutes of Health, Bethesda, Maryland (1991), pp. 647-669 Page), the CDR sequences of 10 mouse monoclonal antibodies were determined (Table 6). Table 5: The amino acid sequence of the variable region of the murine antibody light and heavy chain antibody VH VL SEQ ID NO SEQ ID NO 44F7 1 2 2F12 9 10 3F2 17 18 5F9 25 26 9F3 33 34 10B11 41 42 27B5 49 50 37B1 57 58 44A8 65 66 1D10 73 74 Table 6: Sequence of the CDR of murine antibody antibody VH (SEQ ID NO:) VL (SEQ ID NO:) CDR1 CDR2 CDR3 CDR1 CDR2 CDR3 44F7 3 4 5 6 7 8 2F12 11 12 13 14 15 16 3F2 19 20 twenty one twenty two twenty three twenty four 5F9 27 28 29 30 31 32 9F3 35 36 37 38 39 40 10B11 43 44 45 46 47 48 27B5 51 52 53 54 55 56 37B1 59 60 61 62 63 64 44A8 67 68 69 70 71 72 1D10 75 76 77 78 79 80 3.2 Preparation of human-mouse chimeric antibody and evaluation of antigen binding activity

將編碼上述鼠源抗體的重鏈和輕鏈可變區的基因序列（參見SEQ ID NOs: 103-122）分別與編碼人抗體的重鏈恆定區（SEQ ID NO: 81）和輕鏈恆定區（SEQ ID NO: 82）的序列相連接，並在HEK293細胞（ATCC）中進行重組表達，從而獲得相應的嵌合抗體1D10-chIgG1、2F12-chIgG1、3F2-chIgG1、5F9-chIgG1、9F3-chIgG1、10B11-chIgG1、27B5-chIgG1、37B1-chIgG1、44A8-chIgG1、44F7-chIgG1。並藉由實施例2中所描述的方法檢測不同濃度的嵌合抗體與表達人源CLDN18.2的HEK293T細胞（HEK293T-hCLDN18.2）的結合活性。結果如圖2所示，橫坐標使用抗體濃度的對數，縱坐標使用和CLDN18.2抗體有結合顯示綠色螢光的細胞占貼壁細胞總數的百分比（%螢光細胞）。進一步根據曲線擬合出嵌合抗體結合抗原活性的EC50如表7所示。結果顯示，嵌合抗體3F2-chIgG1、5F9-chIgG1、9F3-chIgG1、10B11-chIgG1、27B5-chIgG1、37B1-chIgG1、44A8-chIgG1、44F7-chIgG1均能夠識別/結合人CLDN18.2。表7：嵌合抗體與CLDN18.2的結合活性測定結果抗體 EC50 (μg/ml) HEK293- hCLDNA18.2 3F2-chIgG1 0.068 5F9-chIgG1 0.283 9F3-chIgG1 0.104 10B11-chIgG1 0.030 27B5-chIgG1 0.108 37B1-chIgG1 0.031 44A8-chIgG1 0.069 44F7-chIgG1 0.027 175D10 0.536 實施例4：抗CLDN18.2嵌合抗體誘導ADCC的活性評價The gene sequences encoding the heavy chain and light chain variable regions of the murine antibody (see SEQ ID NOs: 103-122) were respectively encoded with the heavy chain constant region (SEQ ID NO: 81) and light chain constant region of the human antibody. The sequence of (SEQ ID NO: 82) was connected and recombinantly expressed in HEK293 cells (ATCC) to obtain corresponding chimeric antibodies 1D10-chIgG1, 2F12-chIgG1, 3F2-chIgG1, 5F9-chIgG1, 9F3-chIgG1 , 10B11-chIgG1, 27B5-chIgG1, 37B1-chIgG1, 44A8-chIgG1, 44F7-chIgG1. The method described in Example 2 was used to detect the binding activity of different concentrations of chimeric antibodies to HEK293T cells expressing human CLDN18.2 (HEK293T-hCLDN18.2). The results are shown in Figure 2. The abscissa uses the logarithm of the antibody concentration, and the ordinate uses the percentage of green fluorescent cells (% fluorescent cells) that bind to the CLDN18.2 antibody to the total number of adherent cells. According to the curve fitting, the EC50 of the antigen binding activity of the chimeric antibody is shown in Table 7. The results showed that the chimeric antibodies 3F2-chIgG1, 5F9-chIgG1, 9F3-chIgG1, 10B11-chIgG1, 27B5-chIgG1, 37B1-chIgG1, 44A8-chIgG1, 44F7-chIgG1 were able to recognize/bind human CLDN18.2. Table 7: Results of the determination of the binding activity of the chimeric antibody to CLDN18.2 antibody EC50 (μg/ml) HEK293- hCLDNA18.2 3F2-chIgG1 0.068 5F9-chIgG1 0.283 9F3-chIgG1 0.104 10B11-chIgG1 0.030 27B5-chIgG1 0.108 37B1-chIgG1 0.031 44A8-chIgG1 0.069 44F7-chIgG1 0.027 175D10 0.536 Example 4: Evaluation of anti-CLDN18.2 chimeric antibody inducing ADCC activity

靶細胞使用HEK293T-hCLDN18.2或者天然表達hCLDN18.2的人胃癌腫瘤細胞系KATO-III和NUGC4，效應細胞使用藉由Ficoll分離的人源外周血單核細胞（PBMC）。收穫靶細胞，用PBS清洗2次，將活細胞染料Calcein AM（加入50μl DMSO溶解50μg Calcein AM乾粉，Life Technologies, Cat# C3100MP）稀釋到3μM，在5% CO₂ ，37°C條件下進行靶細胞染色30分鐘。染色完成，用PBS清洗2次，將5000個靶細胞平鋪於100μL DMEM /孔中，使用平底96孔板。藉由將1/3體積(100μL)稀釋於200μL DMEM中來實施3倍梯度稀釋。細胞板的相應孔中加入50μL經稀釋的抗體（非特異性殺傷對照孔加入50μLDMEM培養基）, 在5% CO₂ ，37°C與靶細胞一起孵育30分鐘後，每孔加入50μL分離好的PBMC 50000個作為效應細胞。1000rpm離心3分鐘，讓細胞沉附於板底。在不同時間點0小時，2小時，3小時，4小時，6小時使用全視野細胞掃描分析儀(Nexcelom公司，型號Celigo® Image Cytometer)對實驗板進行測定讀數。測定時選擇Calcein AM對應綠色螢光通道和明場通道同時對孔內細胞進行高速掃描成像，綠色螢光通道得到的成像根據有螢光標記的細胞形態和螢光強度設定參數對孔內的活細胞進行計數，明場通道得到的成像根據細胞形態設定參數對孔內總細胞進行計數，然後兩組資料相除得到顯示綠色螢光的活細胞占細胞總數的百分比。非特異性對照孔的百分比減去抗體的對應百分比得到在該抗體存在的情況下發生特異性細胞裂解的細胞占總細胞的百分比，根據該值判定抗CLDN18.2抗體誘導ADCC的活性，百分比越低代表被測定抗CLDN18.2抗體誘導ADCC的能力越差，反之百分比越高代表被測定抗CLDN18.2抗體誘導ADCC的能力越好。資料分析使用GraphPad。Use HEK293T-hCLDN18.2 or human gastric cancer tumor cell lines KATO-III and NUGC4 that naturally express hCLDN18.2 as target cells, and use human peripheral blood mononuclear cells (PBMC) isolated by Ficoll as effector cells. Harvest the target cells and wash them twice with PBS. Dilute the live cell dye Calcein AM (50μl DMSO to dissolve 50μg Calcein AM dry powder, Life Technologies, Cat# C3100MP) to 3μM, and target under _{5% CO 2, 37°C} The cells were stained for 30 minutes. After staining, wash twice with PBS, and spread 5000 target cells in 100μL DMEM/well, using a flat-bottomed 96-well plate. A 3-fold serial dilution was performed by diluting 1/3 volume (100 μL) in 200 μL DMEM. Add 50μL of diluted antibody to the corresponding wells of the cell plate (add 50μLDMEM medium to the non-specific killing control wells), _{incubate with target cells in 5% CO 2} at 37°C for 30 minutes, then add 50μL of separated PBMC to each well 50000 as effector cells. Centrifuge at 1000 rpm for 3 minutes to allow the cells to settle to the bottom of the plate. At different time points at 0 hour, 2 hours, 3 hours, 4 hours, and 6 hours, the test plate was measured and read using a full-field cell scanning analyzer (Nexcelom company, model Celigo® Image Cytometer). During the measurement, select Calcein AM corresponding to the green fluorescence channel and the bright field channel to simultaneously perform high-speed scanning and imaging of the cells in the well. The cells are counted, and the imaging obtained by the bright field channel counts the total cells in the well according to the cell morphology setting parameters, and then the two sets of data are divided to obtain the percentage of live cells that show green fluorescence to the total number of cells. The percentage of non-specific control wells minus the corresponding percentage of antibody is the percentage of cells that undergo specific cell lysis in the presence of the antibody to the total cells. According to this value, the ADCC-induced activity of the anti-CLDN18.2 antibody is determined. Low represents the poorer the ability of the tested anti-CLDN18.2 antibody to induce ADCC, on the contrary, the higher the percentage represents the better the ability of the tested anti-CLDN18.2 antibody to induce ADCC. Data analysis uses GraphPad.

抗CLDN18.2抗體誘導ADCC活性的測定結果如圖3A-3C所示，橫坐標使用抗體濃度的對數，縱坐標使用校正過的特異性裂解百分比。進一步根據曲線擬合出抗CLDN18.2嵌合抗體誘導ADCC的EC50，其結果如表8所示。結果顯示，所檢測的抗體均可誘導PBMC對於表達人CLDN18.2的細胞的殺傷作用，且明顯優於參比抗體175D10。表8：嵌合抗體誘導ADCC活性的測定結果抗體 ADCC EC50 μg/ml HEK293T-hCLDN18.2 KATO-III NUGC4 3F2-chIgG1 0.0035 5F9-chIgG1 0.0197 9F3-chIgG1 0.0080 10B11-chIgG1 0.0010 27B5-chIgG1 0.0249 37B1-chIgG1 0.0126 44A8-chIgG1 0.0156 44F7-chIgG1 0.0078 0.099 0.047 175D10 0.0580 2.396 N.A. 注：N.A.表示EC50大於測定測定濃度範圍，空白表示該抗體未測量此項資料。實施例5：抗CLDN18.2嵌合抗體誘導CDC的活性評價The results of the determination of the anti-CLDN18.2 antibody-induced ADCC activity are shown in Figures 3A-3C. The abscissa uses the logarithm of the antibody concentration, and the ordinate uses the corrected specific lysis percentage. The EC50 of ADCC induced by the anti-CLDN18.2 chimeric antibody was further calculated according to the curve. The results are shown in Table 8. The results show that the tested antibodies can induce the killing effect of PBMC on the cells expressing human CLDN18.2, and it is significantly better than the reference antibody 175D10. Table 8: Measurement results of chimeric antibody inducing ADCC activity antibody ADCC EC50 μg/ml HEK293T-hCLDN18.2 KATO-III NUGC4 3F2-chIgG1 0.0035 5F9-chIgG1 0.0197 9F3-chIgG1 0.0080 10B11-chIgG1 0.0010 27B5-chIgG1 0.0249 37B1-chIgG1 0.0126 44A8-chIgG1 0.0156 44F7-chIgG1 0.0078 0.099 0.047 175D10 0.0580 2.396 NA Note: NA means that the EC50 is greater than the concentration range of the determination, and blank means that the antibody has not been measured for this data. Example 5: Evaluation of CDC-induced activity of anti-CLDN18.2 chimeric antibody

靶細胞使用HEK293T-hCLDN18.2或者天然表達hCLDN18.2的人胃癌腫瘤細胞系KATO-III，效應細胞使用新鮮人血清。收穫靶細胞，用PBS清洗2次，將活細胞染料Calcein AM（加入50μl DMSO溶解50μg Calcein AM乾粉，Life Technologies, Cat# C3100MP）稀釋到3μM，在5% CO₂ ，37°C條件下進行靶細胞染色30分鐘。染色完成，用PBS清洗2次，將8000個靶細胞平鋪於25μL DMEM /孔中，使用平底96孔板。藉由將1/3體積(100μL)稀釋於200μL DMEM中來實施3倍梯度稀釋。細胞板的每一孔中加入25μL經稀釋的抗體（非特異性殺傷對照孔加入25μLDMEM培養基）, 在5% CO2，37°C與靶細胞一起孵育30分鐘後，每孔加入50μL新鮮分離的20%人血清（用DMEM稀釋）作為效應細胞。1000rpm離心3分鐘，讓細胞沉附於板底。在不同時間點0小時，1小時，2小時，3小時，4小時使用全視野細胞掃描分析儀(Nexcelom公司，型號Celigo® Image Cytometer)對實驗板進行測定讀數。測定時選擇Calcein AM對應綠色螢光通道和明場通道同時對孔內細胞進行高速掃描成像，綠色螢光通道得到的成像根據有螢光標記的細胞形態和螢光強度設定參數對孔內的活細胞進行計數，明場通道得到的成像根據細胞形態設定參數對孔內總細胞進行計數，然後兩組資料相除得顯示綠色螢光的活細胞占細胞總數的百分比，非特異性對照孔的百分比減去抗體的對應百分比得到在該抗體存在的情況下發生特異性細胞殺傷的細胞占總細胞的百分比，根據該值判定抗CLDN18.2抗體誘導CDC的活性，百分比越低代表被測定抗CLDN18.2抗體誘導CDC的能力越差，反之百分比越高代表被測定抗CLDN18.2抗體誘導CDC的能力越好。資料分析使用GraphPad。The target cells used HEK293T-hCLDN18.2 or the human gastric cancer tumor cell line KATO-III that naturally expresses hCLDN18.2, and the effector cells used fresh human serum. Harvest the target cells and wash them twice with PBS. Dilute the live cell dye Calcein AM (50μl DMSO to dissolve 50μg Calcein AM dry powder, Life Technologies, Cat# C3100MP) to 3μM, and target under _{5% CO 2, 37°C} The cells were stained for 30 minutes. After staining is completed, wash twice with PBS, and spread 8000 target cells in 25μL DMEM/well, using a flat-bottomed 96-well plate. A 3-fold serial dilution was performed by diluting 1/3 volume (100 μL) in 200 μL DMEM. Add 25μL of diluted antibody to each well of the cell plate (add 25μLDMEM medium to the non-specific killing control well), incubate with target cells in 5% CO2, 37°C for 30 minutes, and add 50μL of freshly separated 20 to each well % Human serum (diluted with DMEM) is used as effector cells. Centrifuge at 1000 rpm for 3 minutes to allow the cells to settle to the bottom of the plate. At different time points of 0 hour, 1 hour, 2 hours, 3 hours, 4 hours, the whole-field cell scanning analyzer (Nexcelom company, model Celigo® Image Cytometer) was used to measure and read the test plate. During the measurement, select Calcein AM corresponding to the green fluorescence channel and the bright field channel to simultaneously perform high-speed scanning and imaging of the cells in the well. The cells are counted, and the imaging obtained by the bright field channel counts the total cells in the well according to the cell morphology setting parameters, and then the two sets of data are divided to show the percentage of green fluorescent living cells to the total number of cells, and the percentage of non-specific control wells Subtract the corresponding percentage of the antibody to obtain the percentage of cells that have specific cell killing in the presence of the antibody to the total cells. According to this value, the CDC-induced activity of the anti-CLDN18.2 antibody is determined. The lower the percentage, the anti-CLDN18 being determined. 2 The poorer the antibody's ability to induce CDC, conversely, the higher the percentage, the better the ability of the tested anti-CLDN18.2 antibody to induce CDC. Data analysis uses GraphPad.

抗CLDN18.2抗體誘導CDC活性的測定結果如圖4A至圖4B所示，橫坐標使用抗體濃度的對數，縱坐標使用校正過的特異性細胞殺傷百分比。進一步根據曲線擬合出抗CLDN18.2抗體誘導CDC的EC50，其結果如表9所示。結果顯示，所檢測的抗體均可誘導人血清中補體對於表達人CLDN18.2的細胞的殺傷作用，且顯著優於參比抗體175D10。表9：嵌合抗體誘導CDC活性的測定結果抗體 CDC EC50 μg/ml HEK293T-hCLDN18.2 KATO-III 3F2-chIgG1 1.1 5F9-chIgG1 5.25 9F3-chIgG1 1.86 10B11-chIgG1 1.65 27B5-chIgG1 5.17 37B1-chIgG1 1.93 44A8-chIgG1 2.40 44F7-chIgG1 0.60 0.18 175D10 18.18 73.05 注：空白表示該抗體未測量此項資料。實施例6：抗CLDN18.2抗體誘導CLDN18.2內化的活性評價The results of the determination of anti-CLDN18.2 antibody-induced CDC activity are shown in Figure 4A to Figure 4B. The abscissa uses the logarithm of the antibody concentration, and the ordinate uses the corrected specific cell killing percentage. According to the curve fitting, the EC50 of CDC induced by the anti-CLDN18.2 antibody was obtained, and the results are shown in Table 9. The results show that the detected antibodies can all induce the killing effect of complement in human serum on cells expressing human CLDN18.2, and it is significantly better than the reference antibody 175D10. Table 9: Assay results of chimeric antibody inducing CDC activity antibody CDC EC50 μg/ml HEK293T-hCLDN18.2 KATO-III 3F2-chIgG1 1.1 5F9-chIgG1 5.25 9F3-chIgG1 1.86 10B11-chIgG1 1.65 27B5-chIgG1 5.17 37B1-chIgG1 1.93 44A8-chIgG1 2.40 44F7-chIgG1 0.60 0.18 175D10 18.18 73.05 Note: Blank means that the antibody has not been measured for this data. Example 6: Evaluation of the activity of anti-CLDN18.2 antibody to induce CLDN18.2 internalization

本實施例藉由流式細胞術檢測抗CLDN18.2抗體介導細胞（HEK293T-hCLDN18.2）表面的CLDN18.2內化的水準。在37℃下將兩份細胞與10μg/mL嵌合抗體分別孵育1和4小時。用含2% FBS的PBS洗滌若干次後，加入10μg/mL第二抗體4℃下染色30分鐘。然後藉由流式細胞術分析細胞表面CLDN18.2的表達水準。In this example, flow cytometry was used to detect the level of CLDN18.2 internalization on the surface of cells (HEK293T-hCLDN18.2) mediated by the anti-CLDN18.2 antibody. The two cells were incubated with 10 μg/mL chimeric antibody at 37°C for 1 and 4 hours, respectively. After washing several times with PBS containing 2% FBS, 10μg/mL secondary antibody was added for staining at 4°C for 30 minutes. Then the expression level of CLDN18.2 on the cell surface was analyzed by flow cytometry.

MFI_4H 為孵育4小時後的樣品的MFI；MFI_1H 為孵育1小時後的樣品的MFI，假設該條件下抗體完成結合，內吞尚未發生，MFI_背景為僅有第二抗體的MFI，抗體介導的細胞表面CLDN18.2內吞百分百由以下公式計算：內化CLDN18.2的百分比％= 100 - 100 ×(MFI_4H - MFI_1H )/(MFI_1H - MFI_背景 )MFI _4H is the MFI of the sample after 4 hours of incubation; MFI _1H is the MFI of the sample after 1 hour of incubation. It is assumed that the antibody has completed binding under this condition and endocytosis has not yet occurred. The MFI _background is MFI with only the secondary antibody. The percentage of internalization of CLDN18.2 on the cell surface is calculated by the following formula: Percentage of internalization of CLDN18.2% = 100-100 × (MFI _4H -MFI _1H )/(MFI _1H -MFI _background )

結果如表10所示，這些抗體介導了不同程度的HEK293T-hCLDN18.2細胞表面上CLDN18.2的內吞。表10：嵌合抗體誘導CLDN18.2內化的測定結果抗體內吞百分比（％） 1D10-chIgG1 28.7 5F9-chIgG1 9.3 27B5-chIgG1 -2.2 37B1-chIgG1 -3.2 44F7-chIgG1 -1.5 44A8-chIgG1 2.5 實施例7：抗CLDN18.2抗體的人源化及活性評價The results are shown in Table 10. These antibodies mediate the endocytosis of CLDN18.2 on the surface of HEK293T-hCLDN18.2 cells to varying degrees. Table 10: Assay results of the chimeric antibody induced CLDN18.2 internalization antibody Percentage of endocytosis (%) 1D10-chIgG1 28.7 5F9-chIgG1 9.3 27B5-chIgG1 -2.2 37B1-chIgG1 -3.2 44F7-chIgG1 -1.5 44A8-chIgG1 2.5 Example 7: Humanization and activity evaluation of anti-CLDN18.2 antibody

為提高候選抗體與人源抗體的序列的同源性，減少抗體對人的免疫原性，可以對以上實施例提供的鼠源抗體進行人源化設計和製備，使用本領域已知的方法將鼠CDR區***人源框架序列（參見Winter的美國專利No.5,225,539；Queen等人的美國專利Nos.5,530,101; 5,585,089; 5,693,762和6,180,370；以及Lo, Benny, K.C., editor, in Antibody Engineering: Methods and Protocols, volume 248, Humana Press, New Jersey, 2004）。通常而言，人源化抗體的全部或部分CDR區來自於非人源抗體（供體抗體），全部或部分的非CDR區（例如，可變區FR和/或恆定區）來自於人源免疫球蛋白（受體抗體）。In order to improve the sequence homology between the candidate antibody and the human antibody, and reduce the antibody's immunogenicity to humans, the murine antibody provided in the above examples can be humanized design and prepared, using methods known in the art. Mouse CDR regions are inserted into human-derived framework sequences (see U.S. Patent No. 5,225,539 of Winter; U.S. Patent Nos. 5,530,101 of Queen et al.; 5,585,089; 5,693,762 and 6,180,370; and Lo, Benny, KC, editor, in Antibody Engineering: Methods and Protocols , volume 248, Humana Press, New Jersey, 2004). Generally speaking, all or part of the CDR region of a humanized antibody is derived from a non-human antibody (donor antibody), and all or part of the non-CDR region (for example, variable region FR and/or constant region) is derived from human source. Immunoglobulin (receptor antibody).

基於此，本發明人製備獲得了3株鼠源抗體44F7的人源化抗體，分別命名為7004-09hu09、7004-09hu10、7004-09hu15，其氨基酸序列示於下表。表11：人源化抗體的輕重鏈可變區氨基酸序列抗體 7004-09hu09 7004-09hu10 7004-09hu15 SEQ ID NO: VH 99 101 91 HCDR1 75 75 93 HCDR2 76 76 94 HCDR3 77 77 95 VL 100 102 92 LCDR1 78 78 96 LCDR2 79 79 97 LCDR3 80 80 98 Based on this, the present inventors prepared three humanized antibodies of murine antibody 44F7, named 7004-09hu09, 7004-09hu10, 7004-09hu15, and their amino acid sequences are shown in the following table. Table 11: Amino acid sequences of light and heavy chain variable regions of humanized antibodies antibody 7004-09hu09 7004-09hu10 7004-09hu15 SEQ ID NO: VH 99 101 91 HCDR1 75 75 93 HCDR2 76 76 94 HCDR3 77 77 95 VL 100 102 92 LCDR1 78 78 96 LCDR2 79 79 97 LCDR3 80 80 98

將編碼上述人源化抗體的重鏈和輕鏈可變區的基因序列（參見SEQ ID NOs: 123-128）分別與編碼人抗體的重鏈恆定區（SEQ ID NO: 81）和輕鏈恆定區（SEQ ID NO: 82）的序列相連接並進行重組表達。利用實施例2.2中該方法藉由細胞掃描分析儀檢測人源化抗體與HEK293T-hCLDN18.2細胞表面CLDN18.2的結合情況。結果如圖5所示，橫坐標使用抗體濃度的對數，縱坐標使用和CLDN18.2抗體有結合顯示綠色螢光的細胞占貼壁細胞總數的百分比（%螢光細胞）。進一步根據曲線擬合出抗CLDN18.2人源化抗體結合抗原的EC50，結果如表12所示。結果顯示，所檢測的人源化抗體均可結合hCLDN18.2表達的細胞，其親和力和親本嵌合抗體在同一個數量級。表12：人源化抗體與CLDN18.2的結合活性測定結果抗體 EC50 (μg/ml) HEK293- hCLDNA18.2 7004-09hu09 0.051 7004-09hu10 0.047 7004-09hu15 0.008 44F7-chIgG1 0.012 175D10 0.035 The gene sequences encoding the heavy chain and light chain variable regions of the humanized antibody (see SEQ ID NOs: 123-128) were respectively compared with the heavy chain constant region (SEQ ID NO: 81) and light chain constant region of the human antibody. The sequence of the region (SEQ ID NO: 82) is connected and expressed recombinantly. The method in Example 2.2 was used to detect the binding of the humanized antibody to CLDN18.2 on the surface of HEK293T-hCLDN18.2 cells with a cell scanning analyzer. The results are shown in Figure 5. The abscissa uses the logarithm of the antibody concentration, and the ordinate uses the percentage of green fluorescent cells (% fluorescent cells) that bind to CLDN18.2 antibody. The EC50 of the anti-CLDN18.2 humanized antibody binding to the antigen was further calculated according to the curve. The results are shown in Table 12. The results show that the tested humanized antibodies can bind to hCLDN18.2 cells, and their affinity is the same order of magnitude as that of the parental chimeric antibody. Table 12: Results of determination of the binding activity of humanized antibodies to CLDN18.2 antibody EC50 (μg/ml) HEK293- hCLDNA18.2 7004-09hu09 0.051 7004-09hu10 0.047 7004-09hu15 0.008 44F7-chIgG1 0.012 175D10 0.035

進一步，本發明人考察了人源化抗體誘導ADCC的能力，評價方法參見實施例4。靶細胞使用人胃癌腫瘤細胞KATO-III，效應細胞使用藉由Ficoll分離的人源外周血單核細胞。測定結果如圖6所示，橫坐標使用抗體濃度的對數，縱坐標使用校正過的特異性細胞裂解百分比。進一步根據曲線擬合出抗CLDN18.2嵌合抗體誘導ADCC的EC50，其結果如表13所示。結果顯示，所檢測的抗體均可誘導PBMC對於表達人CLDN18.2的細胞的殺傷作用，其誘導ADCC的能力與親本嵌合抗體在同一個數量級，且明顯優於參比抗體175D10。表13：人源化抗體誘導ADCC活性的測定結果抗體 EC50 (μg/ml) KATO-III 7004-09hu09 0.0028 7004-09hu10 0.0033 7004-09hu15 0.0061 44F7-chIgG1 0.0117 175D10 0.7884 Further, the present inventors investigated the ability of humanized antibodies to induce ADCC. For the evaluation method, refer to Example 4. Human gastric cancer tumor cells KATO-III were used as target cells, and human peripheral blood mononuclear cells isolated by Ficoll were used as effector cells. The results of the determination are shown in Figure 6, the abscissa uses the logarithm of the antibody concentration, and the ordinate uses the corrected percentage of specific cell lysis. The EC50 of ADCC induced by the anti-CLDN18.2 chimeric antibody was further calculated according to the curve. The results are shown in Table 13. The results show that the tested antibodies can all induce the killing effect of PBMC on cells expressing human CLDN18.2, and its ADCC inducing ability is the same order of magnitude as that of the parental chimeric antibody, and is significantly better than the reference antibody 175D10. Table 13: Measurement results of humanized antibody-induced ADCC activity antibody EC50 (μg/ml) KATO-III 7004-09hu09 0.0028 7004-09hu10 0.0033 7004-09hu15 0.0061 44F7-chIgG1 0.0117 175D10 0.7884

本發明人還考察了人源化抗體誘導CDC的能力，其評價方法參見實施例5。靶細胞使用人胃癌腫瘤細胞KATO-III，效應細胞使用新鮮人血清。測定結果如圖7所示，橫坐標使用抗體濃度的對數，縱坐標使用校正過的特異性細胞殺傷百分比。進一步根據曲線擬合出抗CLDN18.2嵌合抗體誘導CDC的EC50，其結果如表14所示。結果顯示，所檢測的抗體均可誘導人血清中補體對於表達人CLDN18.2的細胞的殺傷作用，其誘導CDC的能力與親本嵌合抗體在同一個數量級，且明顯優於參比抗體175D10。表14：人源化抗體誘導CDC活性的測定結果抗體 EC50 (μg/ml) KATO-III 7004-09hu09 0.33 7004-09hu10 0.27 7004-09hu15 0.27 44F7-chIgG1 0.52 175D10 34.83 The inventors also investigated the ability of the humanized antibody to induce CDC. Refer to Example 5 for the evaluation method. Human gastric cancer tumor cells KATO-III were used as target cells, and fresh human serum was used as effector cells. The results of the determination are shown in Figure 7, the abscissa uses the logarithm of the antibody concentration, and the ordinate uses the corrected specific cell killing percentage. The EC50 of CDC induced by the anti-CLDN18.2 chimeric antibody was further calculated according to the curve. The results are shown in Table 14. The results show that the tested antibodies can induce the killing effect of complement in human serum on cells expressing human CLDN 18.2, and their CDC-inducing ability is the same order of magnitude as that of the parental chimeric antibody, and is significantly better than the reference antibody 175D10 . Table 14: Determination results of humanized antibody-induced CDC activity antibody EC50 (μg/ml) KATO-III 7004-09hu09 0.33 7004-09hu10 0.27 7004-09hu15 0.27 44F7-chIgG1 0.52 175D10 34.83

以上結果表明，本發明的人源化抗體不僅具有較高的人源化程度，能夠減少免疫排斥反應的可能性，而且展示出與親本嵌合抗體相當且優於已知抗體的抗腫瘤活性。實施例8：抗CLDN18.2嵌合抗體的體內抗腫瘤活性評價The above results indicate that the humanized antibody of the present invention not only has a high degree of humanization and can reduce the possibility of immune rejection, but also exhibits anti-tumor activity comparable to the parental chimeric antibody and superior to known antibodies. . Example 8: Evaluation of in vivo anti-tumor activity of anti-CLDN18.2 chimeric antibody

為了研究抗CLDN18.2抗體在動物體內的抗腫瘤作用，經皮下向裸鼠（SCID, 北京維通利華實驗動物技術有限公司）接種1×10⁷ HEK293T-hCLDN18.2細胞，從接種當天開始進行靜脈內和腹膜內交替給藥，每週兩次，每次10 mg/kg，持續四周，停藥後繼續觀察測量小鼠的腫瘤大小。圖8A顯示了經治療後小鼠腫瘤體積的變化。結果顯示，44F7-chIgG1抗體處理不僅顯著抑制腫瘤的生長，而且完全消除了給藥組小鼠的腫瘤，其抗腫瘤作用顯著優於參比抗體175D10。此外，圖8B顯示44F7-chIgG1顯著延長了荷瘤小鼠的總體生存期，展示了卓越的抗腫瘤活性。由此可見，相比參比抗體175D10，44F7-chIgG1在生存期的延長上也顯示了明顯優勢。這樣的技術效果是顯著的和出人意料的。In order to study the anti-tumor effect of anti-CLDN18.2 antibody in animals, nude mice (SCID, Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd.) were inoculated with 1×10 ⁷ HEK293T-hCLDN18.2 cells subcutaneously, starting from the day of inoculation Alternate intravenous and intraperitoneal administration, twice a week, 10 mg/kg each time for four weeks, continue to observe and measure the tumor size in mice after stopping the drug. Figure 8A shows the change in tumor volume of mice after treatment. The results showed that 44F7-chIgG1 antibody treatment not only significantly inhibited tumor growth, but also completely eliminated tumors in mice in the administration group, and its anti-tumor effect was significantly better than the reference antibody 175D10. In addition, Figure 8B shows that 44F7-chIgG1 significantly prolonged the overall survival of tumor-bearing mice and demonstrated excellent anti-tumor activity. It can be seen that compared with the reference antibody 175D10, 44F7-chIgG1 also shows a significant advantage in prolonging the survival period. This technical effect is remarkable and unexpected.

儘管本發明的具體實施方式已經得到詳細的描述，但本領域技術人員將理解：根據已經公佈的所有教導，可以對細節進行各種修改和變動，並且這些改變均在本發明的保護範圍之內。本發明的全部分為由所附請求項及其任何等同物給出。Although the specific embodiments of the present invention have been described in detail, those skilled in the art will understand that various modifications and changes can be made to the details according to all the teachings that have been published, and these changes are all within the protection scope of the present invention. All the divisions of the present invention are given by the appended claims and any equivalents thereof.

無。none.

圖1A至圖1D分別顯示了抗CLDN18.2鼠源抗體與不同細胞表面CLDN18.2或CLDN18.1的結合活性測定結果。圖1A: HEK293T-hCLDN18.2；圖1B：HEK293T-mCLDN18.2；圖1C：HEK293T-hCLDN18.1；圖1D：HEK293T。圖2顯示了抗CLDN18.2嵌合抗體與細胞表面CLDN18.2的結合活性測定結果。圖3A至圖3C分別顯示了抗CLDN18.2嵌合抗體對HEK293T-hCLDN18.2、KATO-III和NUGC4的ADCC活性的測定結果。圖3A：HEK293T-hCLDN18.2；圖3B: KATO-III；圖3C: NUGC4。圖4A至圖4B分別顯示了抗CLDN18.2嵌合抗體對HEK293T-hCLDN18.2和KATO-III的CDC活性的測定結果。圖4A：HEK293T-hCLDN18.2；圖4B: KATO-III。圖5顯示了抗CLDN18.2人源化抗體與細胞表面CLDN18.2的結合活性測定結果。圖6顯示了抗CLDN18.2人源化抗體對KATO-III的ADCC活性的測定結果。圖7顯示了抗CLDN18.2人源化抗體對KATO-III的CDC活性的測定結果。圖8A至圖8B分別顯示了抗CLDN18.2抗體在小鼠腫瘤模型中對腫瘤體積（A）以及生存期（B）的影響。Figures 1A to 1D respectively show the results of determination of the binding activity of anti-CLDN18.2 murine antibodies to CLDN18.2 or CLDN18.1 on different cell surfaces. Figure 1A: HEK293T-hCLDN18.2; Figure 1B: HEK293T-mCLDN18.2; Figure 1C: HEK293T-hCLDN18.1; Figure 1D: HEK293T. Figure 2 shows the results of the determination of the binding activity of the anti-CLDN18.2 chimeric antibody to the cell surface CLDN18.2. Figures 3A to 3C respectively show the results of the determination of the ADCC activity of the anti-CLDN18.2 chimeric antibody against HEK293T-hCLDN18.2, KATO-III and NUGC4. Figure 3A: HEK293T-hCLDN18.2; Figure 3B: KATO-III; Figure 3C: NUGC4. Figures 4A to 4B respectively show the CDC activity of the anti-CLDN18.2 chimeric antibody on HEK293T-hCLDN18.2 and KATO-III. Figure 4A: HEK293T-hCLDN18.2; Figure 4B: KATO-III. Figure 5 shows the results of the determination of the binding activity of the anti-CLDN18.2 humanized antibody to the cell surface CLDN18.2. Figure 6 shows the results of the determination of the ADCC activity of the anti-CLDN18.2 humanized antibody on KATO-III. Figure 7 shows the results of the determination of the CDC activity of the anti-CLDN18.2 humanized antibody on KATO-III. Figures 8A to 8B respectively show the effect of anti-CLDN18.2 antibody on tumor volume (A) and survival (B) in a mouse tumor model.

Claims

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 3，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 4，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 5，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 6，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 7，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 8，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 3所示的VH CDR1、如SEQ ID NO: 4所示的VH CDR2、如SEQ ID NO: 5所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 6或96所示的VL CDR1、如SEQ ID NO: 7所示的VL CDR2、如SEQ ID NO: 8所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 3, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 4, or has one or several amino acid substitutions, deletions, or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 5, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 6, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 7, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 8, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 3, the VH CDR2 as shown in SEQ ID NO: 4, and the VH CDR3 as shown in SEQ ID NO: 5 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 6 or 96, VL CDR2 as shown in SEQ ID NO: 7 and VL as shown in SEQ ID NO: 8 CDR3.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含一重鏈可變區和一輕鏈可變區，其中， (a) 該重鏈可變區包含SEQ ID NO: 1所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 2所示的輕鏈可變區中含有的3個CDR；或 (b) 該重鏈可變區包含SEQ ID NO: 91所示的重鏈可變區中含有的3個CDR；並且，該輕鏈可變區包含SEQ ID NO: 92所示的輕鏈可變區中含有的3個CDR；較佳地，該重鏈可變區中含有的3個CDR，和/或該輕鏈可變區中含有的3個CDR，由Kabat、Chothia或IMGT編號系統定義。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising a heavy chain variable region and a light chain variable region, wherein, (a) The heavy chain variable region includes the 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 1; and, the light chain variable region includes the light chain variable region shown in SEQ ID NO: 2 3 CDRs contained in the variable region; or (b) The heavy chain variable region includes the 3 CDRs contained in the heavy chain variable region shown in SEQ ID NO: 91; and, the light chain variable region includes the light chain variable region shown in SEQ ID NO: 92 3 CDRs contained in the variable region; Preferably, the 3 CDRs contained in the heavy chain variable region and/or the 3 CDRs contained in the light chain variable region are defined by the Kabat, Chothia or IMGT numbering system.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 75，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 76，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 77，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 78，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 79，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 80，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 75所示的VH CDR1、如SEQ ID NO: 76所示的VH CDR2、如SEQ ID NO: 77所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 78所示的VL CDR1、如SEQ ID NO: 79所示的VL CDR2、如SEQ ID NO: 80所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 75, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 76, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 77, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 78, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 79, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 80, or has one or several amino acid substitutions, deletions, or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 75, the VH CDR2 as shown in SEQ ID NO: 76, and the VH CDR3 as shown in SEQ ID NO: 77 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 78, VL CDR2 as shown in SEQ ID NO: 79, and VL CDR3 as shown in SEQ ID NO: 80.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 11，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 12，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 13，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 14，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 15，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 16，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 11所示的VH CDR1、如SEQ ID NO: 12所示的VH CDR2、如SEQ ID NO: 13所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 14所示的VL CDR1、如SEQ ID NO: 15所示的VL CDR2、如SEQ ID NO: 16所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 11, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 12, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 13, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 14, or compared with it, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 15, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 16, or compared with SEQ ID NO: 16, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 11, the VH CDR2 as shown in SEQ ID NO: 12, and the VH CDR3 as shown in SEQ ID NO: 13 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 14, VL CDR2 as shown in SEQ ID NO: 15 and VL CDR3 as shown in SEQ ID NO: 16.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 19，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 20，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 21，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 22，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 23，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 24，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 19所示的VH CDR1、如SEQ ID NO: 20所示的VH CDR2、如SEQ ID NO: 21所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 22所示的VL CDR1、如SEQ ID NO: 23所示的VL CDR2、如SEQ ID NO: 24所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 19, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 20, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 21, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 22, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 23, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 24, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: VH CDR1 as shown in SEQ ID NO: 19, VH CDR2 as shown in SEQ ID NO: 20, VH CDR3 as shown in SEQ ID NO: 21 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 22, VL CDR2 as shown in SEQ ID NO: 23, and VL CDR3 as shown in SEQ ID NO: 24.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 27，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 28，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 29，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO:30，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 31，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 32，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 27所示的VH CDR1、如SEQ ID NO: 28所示的VH CDR2、如SEQ ID NO: 29所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 30所示的VL CDR1、如SEQ ID NO: 31所示的VL CDR2、如SEQ ID NO: 32所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 27, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 28, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 29, or has one or several amino acid substitutions, deletions, or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 30, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 31, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 32, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 27, the VH CDR2 as shown in SEQ ID NO: 28, and the VH CDR3 as shown in SEQ ID NO: 29 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 30, VL CDR2 as shown in SEQ ID NO: 31, and VL CDR3 as shown in SEQ ID NO: 32.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 35，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 36，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 37，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 38，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 39，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 40，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 35所示的VH CDR1、如SEQ ID NO: 36所示的VH CDR2、如SEQ ID NO: 37所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 38所示的VL CDR1、如SEQ ID NO: 39所示的VL CDR2、如SEQ ID NO: 40所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 35, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 36, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 37, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 38, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 39, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 40, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: VH CDR1 as shown in SEQ ID NO: 35, VH CDR2 as shown in SEQ ID NO: 36, VH CDR3 as shown in SEQ ID NO: 37 ; And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 38, VL CDR2 as shown in SEQ ID NO: 39, and VL CDR3 as shown in SEQ ID NO: 40.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 43，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 44，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 45，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 46，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 47，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 48，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 43所示的VH CDR1、如SEQ ID NO: 44所示的VH CDR2、如SEQ ID NO: 45所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 46所示的VL CDR1、如SEQ ID NO: 47所示的VL CDR2、如SEQ ID NO: 48所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 43, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 44, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 45, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 46, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 47, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 48, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 43, the VH CDR2 as shown in SEQ ID NO: 44, and the VH CDR3 as shown in SEQ ID NO: 45 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 46, VL CDR2 as shown in SEQ ID NO: 47, and VL CDR3 as shown in SEQ ID NO: 48.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 51，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 52，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 53，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 54，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 55，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 56，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 51所示的VH CDR1、如SEQ ID NO: 52所示的VH CDR2、如SEQ ID NO: 53所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 54所示的VL CDR1、如SEQ ID NO: 55所示的VL CDR2、如SEQ ID NO: 56所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 51, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 52, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 53, or compared with it has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 54, or compared with it, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 55, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 56, or compared with it, has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 51, the VH CDR2 as shown in SEQ ID NO: 52, and the VH CDR3 as shown in SEQ ID NO: 53 ; And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 54, VL CDR2 as shown in SEQ ID NO: 55 and VL CDR3 as shown in SEQ ID NO: 56.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 59，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 60，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 61，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 62，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 63，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 64，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 59所示的VH CDR1、如SEQ ID NO: 60所示的VH CDR2、如SEQ ID NO: 61所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 62所示的VL CDR1、如SEQ ID NO: 63所示的VL CDR2、如SEQ ID NO: 64所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 59, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 60, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 61, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 62, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 63, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 64, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: the VH CDR1 as shown in SEQ ID NO: 59, the VH CDR2 as shown in SEQ ID NO: 60, and the VH CDR3 as shown in SEQ ID NO: 61 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 62, VL CDR2 as shown in SEQ ID NO: 63, and VL CDR3 as shown in SEQ ID NO: 64.

一種能夠特異性結合CLDN18.2的抗體或其抗原結合片段，該抗體或其抗原結合片段包含： (a) 包含下述3個互補決定區（CDR）的重鏈可變區（VH）： (i) VH CDR1，其由下述序列組成：SEQ ID NO: 67，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (ii) VH CDR2，其由下述序列組成：SEQ ID NO: 68，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (iii) VH CDR3，其由下述序列組成：SEQ ID NO: 69，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；和/或 (b) 包含下述3個互補決定區（CDR）的輕鏈可變區（VL）： (iv) VL CDR1，其由下述序列組成：SEQ ID NO: 70，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列， (v) VL CDR2，其由下述序列組成：SEQ ID NO: 71，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列，和 (vi) VL CDR3，其由下述序列組成：SEQ ID NO: 72，或與其相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個，2個或3個氨基酸的置換、缺失或添加）的序列；較佳地，(i)-(vi)任一項中所述的置換為保守置換；較佳地，該抗體或其抗原結合片段的VH包含：如SEQ ID NO: 67所示的VH CDR1、如SEQ ID NO: 68所示的VH CDR2、如SEQ ID NO: 69所示的VH CDR3；並且，該抗體或其抗原結合片段的VL包含：如SEQ ID NO: 70所示的VL CDR1、如SEQ ID NO: 71所示的VL CDR2、如SEQ ID NO: 72所示的VL CDR3。An antibody or antigen-binding fragment thereof capable of specifically binding to CLDN18.2, the antibody or antigen-binding fragment thereof comprising: (a) The heavy chain variable region (VH) containing the following 3 complementarity determining regions (CDR): (i) VH CDR1, which consists of the following sequence: SEQ ID NO: 67, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (ii) VH CDR2, which consists of the following sequence: SEQ ID NO: 68, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence, and (iii) VH CDR3, which consists of the following sequence: SEQ ID NO: 69, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2, or 3 amino acid substitutions, deletions) Or add) sequence; and / or (b) The light chain variable region (VL) containing the following 3 complementarity determining regions (CDR): (iv) VL CDR1, which consists of the following sequence: SEQ ID NO: 70, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, (v) VL CDR2, which consists of the following sequence: SEQ ID NO: 71, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence, and (vi) VL CDR3, which consists of the following sequence: SEQ ID NO: 72, or has one or several amino acid substitutions, deletions or additions (for example, 1, 2 or 3 amino acid substitutions, deletions) Or add) sequence; Preferably, the substitution described in any one of (i)-(vi) is a conservative substitution; Preferably, the VH of the antibody or antigen-binding fragment thereof comprises: VH CDR1 as shown in SEQ ID NO: 67, VH CDR2 as shown in SEQ ID NO: 68, VH CDR3 as shown in SEQ ID NO: 69 And, the VL of the antibody or antigen-binding fragment thereof comprises: VL CDR1 as shown in SEQ ID NO: 70, VL CDR2 as shown in SEQ ID NO: 71, and VL CDR3 as shown in SEQ ID NO: 72.

如請求項1或請求項2所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 1所示的序列； (ii) 與SEQ ID NO: 1所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 1所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 2所示的序列； (v) 與SEQ ID NO: 2所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 2所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 1所示的序列的VH和具有如SEQ ID NO:2所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 1 or claim 2, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 1; (ii) Compared with the sequence shown in SEQ ID NO: 1, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 1 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 2; (v) Compared with the sequence shown in SEQ ID NO: 2, there are one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 2 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 1 and VL having the sequence shown in SEQ ID NO: 2.

如請求項1至請求項11中任一項所述的抗體或其抗原結合片段，其中該抗體是人源化的。The antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 11, wherein the antibody is humanized.

請求項2或請求項13所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 91所示的序列； (ii) 與SEQ ID NO: 91所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 91所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 92所示的序列； (v) 與SEQ ID NO: 92所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 92所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 91所示的序列的VH和具有如SEQ ID NO:92所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 2 or claim 13, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 91; (ii) Compared with the sequence shown in SEQ ID NO: 91, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 91 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 92; (v) Compared with the sequence shown in SEQ ID NO: 92, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 92 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 91 and VL having the sequence shown in SEQ ID NO: 92.

請求項2或請求項13所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 99所示的序列； (ii) 與SEQ ID NO: 99所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 99所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 100所示的序列； (v) 與SEQ ID NO: 100所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 100所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 99所示的序列的VH和具有如SEQ ID NO: 100所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 2 or claim 13, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 99; (ii) Compared with the sequence shown in SEQ ID NO: 99, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 99 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 100; (v) Compared with the sequence shown in SEQ ID NO: 100, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 100 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 99 and VL having the sequence shown in SEQ ID NO: 100.

請求項2或請求項13所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 101所示的序列； (ii) 與SEQ ID NO: 101所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 101所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 102所示的序列； (v) 與SEQ ID NO: 102所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 102所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 101所示的序列的VH和具有如SEQ ID NO: 102所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 2 or claim 13, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 101; (ii) Compared with the sequence shown in SEQ ID NO: 101, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 101 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 102; (v) Compared with the sequence shown in SEQ ID NO: 102, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 102 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 101 and VL having the sequence shown in SEQ ID NO: 102.

如請求項3所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 73所示的序列； (ii) 與SEQ ID NO: 73所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 73所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 74所示的序列； (v) 與SEQ ID NO: 74所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 74所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 73所示的序列的VH和具有如SEQ ID NO: 74所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 3, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 73; (ii) Compared with the sequence shown in SEQ ID NO: 73, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 73 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 74; (v) Compared with the sequence shown in SEQ ID NO: 74, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 74 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 73 and VL having the sequence shown in SEQ ID NO: 74.

如請求項4所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 9所示的序列； (ii) 與SEQ ID NO: 9所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 9所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 10所示的序列； (v) 與SEQ ID NO: 10所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 10所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 9所示的序列的VH和具有如SEQ ID NO: 10所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 4, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 9; (ii) Compared with the sequence shown in SEQ ID NO: 9 has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions ) Sequence; or (iii) The sequence shown in SEQ ID NO: 9 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 10; (v) Compared with the sequence shown in SEQ ID NO: 10, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 10 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 9 and VL having the sequence shown in SEQ ID NO: 10.

如請求項5所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 17所示的序列； (ii) 與SEQ ID NO: 17所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 17所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 18所示的序列； (v) 與SEQ ID NO: 18所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 18所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 17所示的序列的VH和具有如SEQ ID NO: 18所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 5, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 17; (ii) Compared with the sequence shown in SEQ ID NO: 17 has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions ) Sequence; or (iii) The sequence shown in SEQ ID NO: 17 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 18; (v) Compared with the sequence shown in SEQ ID NO: 18, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 18 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 17 and VL having the sequence shown in SEQ ID NO: 18.

如請求項6所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 25所示的序列； (ii) 與SEQ ID NO: 25所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 25所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 26所示的序列； (v) 與SEQ ID NO: 26所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 26所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 25所示的序列的VH和具有如SEQ ID NO: 26所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 6, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 25; (ii) Compared with the sequence shown in SEQ ID NO: 25, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 25 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 26; (v) Compared with the sequence shown in SEQ ID NO: 26, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 26 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 25 and VL having the sequence shown in SEQ ID NO: 26.

如請求項7所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 33所示的序列； (ii) 與SEQ ID NO: 33所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 33所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 34所示的序列； (v) 與SEQ ID NO: 34所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 34所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 33所示的序列的VH和具有如SEQ ID NO: 34所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 7, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 33; (ii) Compared with the sequence shown in SEQ ID NO: 33, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 33 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 34; (v) Compared with the sequence shown in SEQ ID NO: 34, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 34 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 33 and VL having the sequence shown in SEQ ID NO: 34.

如請求項8所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 41所示的序列； (ii) 與SEQ ID NO: 41所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 41所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 42所示的序列； (v) 與SEQ ID NO: 42所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 42所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 41所示的序列的VH和具有如SEQ ID NO: 42所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 8, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 41; (ii) Compared with the sequence shown in SEQ ID NO: 41, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 41 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 42; (v) Compared with the sequence shown in SEQ ID NO: 42 there are one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions ) Sequence; or (vi) The sequence shown in SEQ ID NO: 42 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 41 and VL having the sequence shown in SEQ ID NO: 42.

如請求項9所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 49所示的序列； (ii) 與SEQ ID NO: 49所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 49所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 50所示的序列； (v) 與SEQ ID NO: 50所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 50所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 49所示的序列的VH和具有如SEQ ID NO: 50所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 9, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 49; (ii) Compared with the sequence shown in SEQ ID NO: 49, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 49 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 50; (v) Compared with the sequence shown in SEQ ID NO: 50, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 50 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 49 and VL having the sequence shown in SEQ ID NO: 50.

如請求項10所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 57所示的序列； (ii) 與SEQ ID NO: 57所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 57所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 58所示的序列； (v) 與SEQ ID NO: 58所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 58所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 57所示的序列的VH和具有如SEQ ID NO: 58所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 10, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 57; (ii) Compared with the sequence shown in SEQ ID NO: 57, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 57 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 58; (v) Compared with the sequence shown in SEQ ID NO: 58 there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 58 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 57 and VL having the sequence shown in SEQ ID NO: 58.

如請求項11所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段包含： (a) 重鏈可變區（VH），其包含選自下列的氨基酸序列： (i) SEQ ID NO: 65所示的序列； (ii) 與SEQ ID NO: 65所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (iii) 與SEQ ID NO: 65所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；和/或， (b) 輕鏈可變區（VL），其包含選自下列的氨基酸序列： (iv) SEQ ID NO: 66所示的序列； (v) 與SEQ ID NO: 66所示的序列相比具有一個或幾個氨基酸的置換、缺失或添加（例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）的序列；或 (vi) 與SEQ ID NO: 66所示的序列具有至少80%、至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、至少99%、或100%的序列同一性的序列；較佳地，(ii)或(v)中所述的置換是保守置換；較佳地，該抗體或其抗原結合片段包含：具有如SEQ ID NO: 65所示的序列的VH和具有如SEQ ID NO: 66所示的序列的VL。The antibody or antigen-binding fragment thereof according to claim 11, wherein the antibody or antigen-binding fragment thereof comprises: (a) The variable region of the heavy chain (VH), which comprises an amino acid sequence selected from the following: (i) The sequence shown in SEQ ID NO: 65; (ii) Compared with the sequence shown in SEQ ID NO: 65, it has one or several amino acid substitutions, deletions or additions (for example, 1, 2, 3, 4 or 5 amino acid substitutions, deletions or additions) ) Sequence; or (iii) The sequence shown in SEQ ID NO: 65 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; and / or, (b) The light chain variable region (VL), which comprises an amino acid sequence selected from the following: (iv) The sequence shown in SEQ ID NO: 66; (v) Compared with the sequence shown in SEQ ID NO: 66, there are substitutions, deletions or additions of one or several amino acids (for example, substitutions, deletions or additions of 1, 2, 3, 4 or 5 amino acids) ) Sequence; or (vi) The sequence shown in SEQ ID NO: 66 has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, A sequence with at least 97%, at least 98%, at least 99%, or 100% sequence identity; Preferably, the substitutions described in (ii) or (v) are conservative substitutions; Preferably, the antibody or antigen-binding fragment thereof comprises: VH having the sequence shown in SEQ ID NO: 65 and VL having the sequence shown in SEQ ID NO: 66.

如請求項1至請求項25中任一項所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段進一步包含： (a) 人免疫球蛋白的重鏈恆定區（CH）或其變體，該變體與其所源自的序列相比具有一或複數氨基酸的置換、缺失或添加（例如，至多20個、至多15個、至多10個、或至多5個氨基酸的置換、缺失或添加；例如1個、2個、3個、4個或5個氨基酸的置換、缺失或添加）；和 (b) 人免疫球蛋白的輕鏈恆定區（CL）或其變體，該變體與其所源自的序列相比具有至多20個氨基酸的保守置換（例如至多15個、至多10個、或至多5個氨基酸的保守置換；例如1個、2個、3個、4個或5個氨基酸的保守置換）；較佳地，該重鏈恆定區是IgG重鏈恆定區，例如IgG1、IgG2、IgG3或IgG4重鏈恆定區；較佳地，該抗體或其抗原結合片段包含SEQ ID NO: 81所示的重鏈恆定區（CH）；較佳地，該輕鏈恆定區是κ輕鏈恆定區；較佳地，該抗體或其抗原結合片段包含SEQ ID NO: 82所示的輕鏈恆定區（CL）。The antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 25, wherein the antibody or antigen-binding fragment thereof further comprises: (a) The heavy chain constant region (CH) of a human immunoglobulin or a variant thereof, which has one or more amino acid substitutions, deletions or additions (for example, at most 20, at most 15, at most 10, or at most 5 amino acid substitutions, deletions, or additions; for example, 1, 2, 3, 4, or 5 amino acid substitutions, deletions, or additions); and (b) The light chain constant region (CL) of a human immunoglobulin or a variant thereof, which has conservative substitutions of at most 20 amino acids (for example, at most 15, at most 10, or at most) compared with the sequence from which it is derived Conservative substitutions of up to 5 amino acids; for example, conservative substitutions of 1, 2, 3, 4, or 5 amino acids); Preferably, the heavy chain constant region is an IgG heavy chain constant region, such as an IgG1, IgG2, IgG3 or IgG4 heavy chain constant region; Preferably, the antibody or antigen-binding fragment thereof comprises the heavy chain constant region (CH) shown in SEQ ID NO: 81; Preferably, the light chain constant region is a kappa light chain constant region; Preferably, the antibody or antigen-binding fragment thereof comprises a light chain constant region (CL) shown in SEQ ID NO: 82.

如請求項1至請求項26中任一項所述的抗體或其抗原結合片段，其中，該抗原結合片段選自Fab、Fab’、(Fab’)₂ 、Fv、二硫鍵連接的Fv、scFv、雙抗體(diabody)和單域抗體（sdAb）；和/或，該抗體為鼠源抗體、嵌合抗體、人源化抗體、雙特異性抗體或多特異性抗體。The antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 26, wherein the antigen-binding fragment is selected from Fab, Fab', (Fab') ₂ , Fv, disulfide-linked Fv, scFv, diabody and single domain antibody (sdAb); and/or, the antibody is a murine antibody, a chimeric antibody, a humanized antibody, a bispecific antibody or a multispecific antibody.

如請求項1至請求項27中任一項所述的抗體或其抗原結合片段，其中，該抗體或其抗原結合片段帶有標記；較佳地，該抗體或其抗原結合片段帶有可檢測的標記，例如酶（例如辣根過氧化物酶）、放射性核素、螢光染料、發光物質（如化學發光物質）或生物素。The antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 27, wherein the antibody or antigen-binding fragment thereof bears a label; preferably, the antibody or antigen-binding fragment thereof bears a detectable Labels such as enzymes (such as horseradish peroxidase), radionuclides, fluorescent dyes, luminescent substances (such as chemiluminescent substances) or biotin.

一種分離的核酸分子，其編碼如請求項1至請求項28中任一項所述的抗體或其抗原結合片段，或其重鏈可變區和/或輕鏈可變區。An isolated nucleic acid molecule that encodes the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28, or its heavy chain variable region and/or light chain variable region.

一種載體，其包含如請求項29所述的分離的核酸分子；較佳地，該載體為殖株載體或表達載體。A vector comprising the isolated nucleic acid molecule as described in claim 29; preferably, the vector is a clone vector or an expression vector.

一種宿主細胞，其包含如請求項29所述的分離的核酸分子或請求項30所述的載體。A host cell comprising the isolated nucleic acid molecule according to claim 29 or the vector according to claim 30.

一種製備如請求項1至請求項28中任一項所述的抗體或其抗原結合片段的方法，其包括，在允許該抗體或其抗原結合片段表達的條件下，培養請求項31所述的宿主細胞，和從培養的宿主細胞培養物中回收該抗體或其抗原結合片段。A method for preparing the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28, which comprises culturing the antibody or antigen-binding fragment thereof under conditions that allow expression of the antibody or antigen-binding fragment thereof. Host cells, and recovering the antibody or antigen-binding fragment thereof from the cultured host cell culture.

一種雙特異性或多特異性分子，其包含如請求項1至請求項28中任一項所述的抗體或其抗原結合片段；較佳地，該雙特異性或多特異性分子特異性結合CLDN18.2，並且額外地特異性結合一或複數其他靶標；較佳地，該雙特異性或多特異性分子還包含至少一種具有針對第二靶標的第二結合特異性的分子（例如第二抗體）。A bispecific or multispecific molecule comprising the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28; Preferably, the bispecific or multispecific molecule specifically binds to CLDN 18.2, and additionally specifically binds to one or more other targets; Preferably, the bispecific or multispecific molecule further comprises at least one molecule (such as a second antibody) having a second binding specificity for a second target.

一種免疫綴合物，其包含如請求項1至請求項28中任一項所述的抗體或其抗原結合片段以及連接於該抗體或其抗原結合片段的一治療劑；較佳地，該治療劑選自細胞毒劑；較佳地，該治療劑選自烷化劑、有絲***抑制劑、抗腫瘤抗生素、抗代謝物、拓撲異構酶抑制劑、酪氨酸激酶抑制劑、放射性核素劑，及其任意組合；較佳地，該免疫綴合物是抗體-藥物偶聯物（ADC）。An immunoconjugate comprising the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28, and a therapeutic agent linked to the antibody or antigen-binding fragment thereof; Preferably, the therapeutic agent is selected from cytotoxic agents; Preferably, the therapeutic agent is selected from alkylating agents, mitotic inhibitors, anti-tumor antibiotics, antimetabolites, topoisomerase inhibitors, tyrosine kinase inhibitors, radionuclide agents, and any combination thereof; Preferably, the immunoconjugate is an antibody-drug conjugate (ADC).

一種藥物組合物，其含有如請求項1至請求項28中任一項所述的抗體或其抗原結合片段、請求項33所述的雙特異性或多特異性分子或者請求項34所述的免疫綴合物，以及藥學上可接受的載體和/或賦形劑；較佳地，藥物組合物還包含另外的藥學活性劑；較佳地，該另外的藥學活性劑是具有抗腫瘤活性的藥物，例如烷化劑、有絲***抑制劑、抗腫瘤抗生素、抗代謝物、拓撲異構酶抑制劑、酪氨酸激酶抑制劑、放射性核素劑、放射增敏劑、抗血管產生劑、細胞因數、分子靶向藥物、免疫檢查點抑制劑或溶瘤病毒；較佳地，該抗體或其抗原結合片段、雙特異性或多特異性分子或免疫綴合物與所述另外的藥學活性劑作為分離的組分或作為同一組合物的組分提供。A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28, the bispecific or multispecific molecule according to claim 33, or the antibody described in claim 34 Immunoconjugates, and pharmaceutically acceptable carriers and/or excipients; Preferably, the pharmaceutical composition further comprises another pharmaceutically active agent; Preferably, the additional pharmacologically active agent is a drug with anti-tumor activity, such as alkylating agents, mitotic inhibitors, anti-tumor antibiotics, antimetabolites, topoisomerase inhibitors, tyrosine kinase inhibitors, radioactive Nuclide agents, radiosensitizers, anti-angiogenic agents, cytokines, molecular targeted drugs, immune checkpoint inhibitors or oncolytic viruses; Preferably, the antibody or antigen-binding fragment, bispecific or multispecific molecule or immunoconjugate and the additional pharmaceutically active agent are provided as separate components or as components of the same composition.

一種試劑盒，其含有如請求項1至請求項28中任一項所述的抗體或其抗原結合片段；較佳地，該抗體或其抗原結合片段帶有可檢測的標記，例如酶（例如辣根過氧化物酶）、放射性核素、螢光染料、發光物質（如化學發光物質）或生物素；較佳地，該試劑盒還包括一第二抗體，其特異性識別請求項1至請求項28中任一項所述的抗體或其抗原結合片段；較佳地，該第二抗體還包括可檢測的標記，例如酶（例如辣根過氧化物酶）、放射性核素、螢光染料、發光物質（如化學發光物質）或生物素。A kit containing the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28; Preferably, the antibody or antigen-binding fragment thereof has a detectable label, such as an enzyme (such as horseradish peroxidase), a radionuclide, a fluorescent dye, a luminescent substance (such as a chemiluminescent substance) or biotin; Preferably, the kit further includes a second antibody, which specifically recognizes the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28; Preferably, the second antibody also includes a detectable label, such as an enzyme (such as horseradish peroxidase), a radionuclide, a fluorescent dye, a luminescent substance (such as a chemiluminescent substance) or biotin.

一種嵌合抗原受體，其包含如請求項1至請求項28中任一項所述的抗體或其抗原結合片段的抗原結合結構域；較佳地，該抗原結合結構域包含如請求項1至請求項28中任一項所述的抗體或其抗原結合片段的重鏈可變區和輕鏈可變區；較佳地，該抗原結合結構域是scFv；較佳地，該嵌合抗原受體包含如請求項1至請求項28中任一項所述的抗體的抗原結合片段；較佳地，該嵌合抗原受體由免疫效應細胞（例如T細胞）所表達。A chimeric antigen receptor comprising the antigen-binding domain of the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28; Preferably, the antigen-binding domain comprises the heavy chain variable region and the light chain variable region of the antibody or antigen-binding fragment thereof as described in any one of claim 1 to claim 28; Preferably, the antigen binding domain is scFv; Preferably, the chimeric antigen receptor comprises an antigen-binding fragment of the antibody according to any one of claim 1 to claim 28; Preferably, the chimeric antigen receptor is expressed by immune effector cells (such as T cells).

一種分離的核酸分子，其編碼如請求項37所述的嵌合抗原受體。An isolated nucleic acid molecule encoding the chimeric antigen receptor according to claim 37.

一種載體，其包含如請求項38所述的分離的核酸分子；較佳地，其用於製備嵌合抗原受體T細胞。A vector comprising the isolated nucleic acid molecule as described in claim 38; preferably, it is used to prepare chimeric antigen receptor T cells.

一種宿主細胞，其包含如請求項38所述的分離的核酸分子或請求項39所述的載體；較佳地，該宿主細胞是免疫效應細胞（例如，T細胞或NK細胞）；較佳地，該宿主細胞是嵌合抗原受體T細胞（CAR-T）。A host cell comprising the isolated nucleic acid molecule according to claim 38 or the vector according to claim 39; Preferably, the host cell is an immune effector cell (for example, T cell or NK cell); Preferably, the host cell is a chimeric antigen receptor T cell (CAR-T).

一種用於降低CLDN18.2在細胞表面的表達水準的方法，其包括將該細胞與請求項1至請求項28中任一項所述的抗體或其抗原結合片段，或請求項33所述的雙特異性或多特異性分子，或請求項34所述的免疫綴合物，或請求項35所述的藥物組合物，或請求項37所述的嵌合抗原受體，或請求項40所述的宿主細胞接觸，使得CLDN18.2在該細胞表面的表達水準降低；其中，該細胞在其表面表達CLDN18.2；較佳地，該細胞是表達CLDN18.2的腫瘤細胞。A method for reducing the expression level of CLDN18.2 on the cell surface, which comprises combining the cell with the antibody or antigen-binding fragment thereof described in any one of claim 1 to claim 28, or the method described in claim 33 Bispecific or multispecific molecule, or immunoconjugate according to claim 34, or pharmaceutical composition according to claim 35, or chimeric antigen receptor according to claim 37, or claim 40 The host cell contact described above reduces the expression level of CLDN18.2 on the cell surface; wherein, the cell expresses CLDN18.2 on its surface; Preferably, the cell is a tumor cell expressing CLDN 18.2.

一種抑制表達CLDN18.2的腫瘤細胞生長和/或殺傷該腫瘤細胞的方法，其包括將該腫瘤細胞與有效量的請求項1至請求項28中任一項所述的抗體或其抗原結合片段，或請求項33所述的雙特異性或多特異性分子，或請求項34所述的免疫綴合物，或請求項35所述的藥物組合物，或請求項37所述的嵌合抗原受體，或請求項40所述的宿主細胞接觸。A method for inhibiting the growth of tumor cells expressing CLDN18.2 and/or killing the tumor cells, which comprises combining the tumor cells with an effective amount of the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28 , Or the bispecific or multispecific molecule of claim 33, or the immunoconjugate of claim 34, or the pharmaceutical composition of claim 35, or the chimeric antigen of claim 37 Recipient, or contact with the host cell described in claim 40.

一種用於在一受試者（例如人）中預防和/或治療腫瘤的方法，該方法包括向有此需要的受試者施用有效量的請求項1至請求項28中任一項所述的抗體或其抗原結合片段，或請求項33所述的雙特異性或多特異性分子，或請求項34所述的免疫綴合物，或請求項35所述的藥物組合物，或請求項37所述的嵌合抗原受體，或請求項40所述的宿主細胞；較佳地，該腫瘤表達CLDN18.2；較佳地，該腫瘤涉及表達CLDN18.2的腫瘤細胞；較佳地，該CLDN18.2在該腫瘤細胞表面上表達；較佳地，該腫瘤選自胃癌、食道癌、胰腺癌、支氣管癌、非小細胞肺癌、乳腺癌、耳鼻喉(ENT)癌、卵巢癌、結腸癌、肝癌、頭頸癌、膽囊癌及其轉移癌（例如，胃癌轉移，如Krukenberg瘤、腹膜轉移或淋巴結轉移）；較佳地，該受試者為哺乳動物，例如人；較佳地，該方法還包括施用另外的具有抗腫瘤活性的藥物，例如烷化劑、有絲***抑制劑、抗腫瘤抗生素、抗代謝物、拓撲異構酶抑制劑、酪氨酸激酶抑制劑、放射性核素劑、放射增敏劑、抗血管產生劑、細胞因數、分子靶向藥物、免疫檢查點抑制劑或溶瘤病毒；較佳地，該方法還包括施用另外的抗腫瘤療法，例如手術、化學治療、放射治療、靶向治療、免疫治療、激素治療、基因治療或姑息治療。A method for preventing and/or treating tumors in a subject (such as a human), the method comprising administering an effective amount of any one of claim 1 to claim 28 to a subject in need The antibody or antigen-binding fragment thereof, or the bispecific or multispecific molecule according to claim 33, or the immunoconjugate according to claim 34, or the pharmaceutical composition according to claim 35, or The chimeric antigen receptor according to 37, or the host cell according to claim 40; Preferably, the tumor expresses CLDN 18.2; Preferably, the tumor involves tumor cells expressing CLDN 18.2; preferably, the CLDN 18.2 is expressed on the surface of the tumor cells; Preferably, the tumor is selected from gastric cancer, esophageal cancer, pancreatic cancer, bronchial cancer, non-small cell lung cancer, breast cancer, ear, nose and throat (ENT) cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer, gallbladder cancer and its metastasis Cancer (for example, gastric cancer metastasis, such as Krukenberg tumor, peritoneal metastasis, or lymph node metastasis); Preferably, the subject is a mammal, such as a human; Preferably, the method also includes the administration of other drugs with anti-tumor activity, such as alkylating agents, mitotic inhibitors, anti-tumor antibiotics, antimetabolites, topoisomerase inhibitors, tyrosine kinase inhibitors, radioactive Nuclide agents, radiosensitizers, anti-angiogenic agents, cytokines, molecular targeted drugs, immune checkpoint inhibitors or oncolytic viruses; Preferably, the method also includes the administration of additional anti-tumor therapy, such as surgery, chemotherapy, radiation therapy, targeted therapy, immunotherapy, hormone therapy, gene therapy or palliative therapy.

如請求項1至請求項28中任一項所述的抗體或其抗原結合片段，或請求項33所述的雙特異性或多特異性分子，或請求項34所述的免疫綴合物，或請求項35所述的藥物組合物，或請求項37所述的嵌合抗原受體，或請求項38所述的分離的核酸分子，或請求項39所述的載體，或請求項40所述的宿主細胞，在製備藥物中的用途，該藥物用於在受試者（例如人）中預防和/治療腫瘤；較佳地，藥物還包含一另外的藥學活性劑；較佳地，所述另外的藥學活性劑是具有抗腫瘤活性的藥物，例如烷化劑、有絲***抑制劑、抗腫瘤抗生素、抗代謝物、拓撲異構酶抑制劑、酪氨酸激酶抑制劑、放射性核素劑、放射增敏劑、抗血管產生劑、細胞因數、分子靶向藥物、免疫檢查點抑制劑或溶瘤病毒；較佳地，該腫瘤表達CLDN18.2；較佳地，該腫瘤涉及表達CLDN18.2的腫瘤細胞；較佳地，該CLDN18.2在該腫瘤細胞表面上表達；較佳地，該腫瘤選自胃癌、食道癌、胰腺癌、支氣管癌、非小細胞肺癌、乳腺癌、耳鼻喉(ENT)癌、卵巢癌、結腸癌、肝癌、頭頸癌、膽囊癌及其轉移癌（例如，胃癌轉移，如Krukenberg瘤、腹膜轉移或淋巴結轉移）；較佳地，該受試者為哺乳動物，例如人。The antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28, or the bispecific or multispecific molecule according to claim 33, or the immunoconjugate according to claim 34, Or the pharmaceutical composition according to claim 35, or the chimeric antigen receptor according to claim 37, or the isolated nucleic acid molecule according to claim 38, or the vector according to claim 39, or the chimeric antigen receptor according to claim 40 The use of the host cell in the preparation of a medicament for the prevention and/treatment of tumors in subjects (such as humans); Preferably, the medicine also contains an additional pharmaceutically active agent; Preferably, the additional pharmacologically active agent is a drug with anti-tumor activity, such as alkylating agents, mitotic inhibitors, anti-tumor antibiotics, antimetabolites, topoisomerase inhibitors, tyrosine kinase inhibitors, Radionuclide agents, radiosensitizers, anti-angiogenic agents, cytokines, molecular targeted drugs, immune checkpoint inhibitors or oncolytic viruses; Preferably, the tumor expresses CLDN 18.2; Preferably, the tumor involves tumor cells expressing CLDN 18.2; preferably, the CLDN 18.2 is expressed on the surface of the tumor cells; Preferably, the tumor is selected from gastric cancer, esophageal cancer, pancreatic cancer, bronchial cancer, non-small cell lung cancer, breast cancer, ear, nose and throat (ENT) cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer, gallbladder cancer and its metastasis Cancer (for example, gastric cancer metastasis, such as Krukenberg tumor, peritoneal metastasis, or lymph node metastasis); Preferably, the subject is a mammal, such as a human.

一種檢測CLDN18.2（例如人CLDN18.2）在一樣品中的存在或其量的方法，其包括以下步驟： (1) 將該樣品與請求項1至請求項28中任一項所述的抗體或其抗原結合片段接觸； (2) 檢測該抗體或其抗原結合片段與CLDN18.2之間複合物的形成或檢測該複合物的量；較佳地，該抗體或其抗原結合片段帶有可檢測的標記；較佳地，該CLDN18.2是人CLDN18.2。A method for detecting the presence or amount of CLDN 18.2 (such as human CLDN 18.2) in a sample, which includes the following steps: (1) Contact the sample with the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28; (2) Detect the formation of the complex between the antibody or its antigen-binding fragment and CLDN18.2 or detect the amount of the complex; Preferably, the antibody or antigen-binding fragment thereof has a detectable label; Preferably, the CLDN 18.2 is human CLDN 18.2.

一種用於檢測一腫瘤是否能夠藉由靶向CLDN18.2的抗腫瘤療法來治療的方法，其包括以下步驟： (1) 將含有該腫瘤細胞的樣品與請求項1至請求項28中任一項所述的抗體或其抗原結合片段接觸； (2) 檢測該抗體或其抗原結合片段與CLDN18.2之間複合物的形成；較佳地，該抗體或其抗原結合片段帶有可檢測的標記；較佳地，該CLDN18.2是人CLDN18.2；較佳地，該腫瘤選自胃癌、食道癌、胰腺癌、支氣管癌、非小細胞肺癌、乳腺癌、耳鼻喉(ENT)癌、卵巢癌、結腸癌、肝癌、頭頸癌、膽囊癌及其轉移癌（例如，胃癌轉移，如Krukenberg瘤、腹膜轉移或淋巴結轉移）。A method for detecting whether a tumor can be treated by anti-tumor therapy targeting CLDN 18.2, which includes the following steps: (1) Contacting a sample containing the tumor cell with the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28; (2) Detect the formation of the complex between the antibody or its antigen-binding fragment and CLDN18.2; Preferably, the antibody or antigen-binding fragment thereof has a detectable label; Preferably, the CLDN 18.2 is human CLDN 18.2; Preferably, the tumor is selected from gastric cancer, esophageal cancer, pancreatic cancer, bronchial cancer, non-small cell lung cancer, breast cancer, ear, nose and throat (ENT) cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer, gallbladder cancer and its metastasis Cancer (for example, gastric cancer metastasis, such as Krukenberg tumor, peritoneal metastasis, or lymph node metastasis).

請求項1至請求項28中任一項所述的抗體或其抗原結合片段在製備一試劑盒中的用途，該試劑盒用於檢測腫瘤是否能夠藉由靶向CLDN18.2的抗腫瘤療法來治療；較佳地，該抗體或其抗原結合片段帶有可檢測的標記；較佳地，該CLDN18.2是人CLDN18.2；較佳地，該腫瘤選自胃癌、食道癌、胰腺癌、支氣管癌、非小細胞肺癌、乳腺癌、耳鼻喉(ENT)癌、卵巢癌、結腸癌、肝癌、頭頸癌、膽囊癌及其轉移癌（例如，胃癌轉移，如Krukenberg瘤、腹膜轉移或淋巴結轉移）。Use of the antibody or antigen-binding fragment thereof according to any one of claim 1 to claim 28 in the preparation of a kit for detecting whether a tumor can be treated by anti-tumor therapy targeting CLDN 18.2 treat; Preferably, the antibody or antigen-binding fragment thereof has a detectable label; Preferably, the CLDN 18.2 is human CLDN 18.2; Preferably, the tumor is selected from gastric cancer, esophageal cancer, pancreatic cancer, bronchial cancer, non-small cell lung cancer, breast cancer, ear, nose and throat (ENT) cancer, ovarian cancer, colon cancer, liver cancer, head and neck cancer, gallbladder cancer and its metastasis Cancer (for example, gastric cancer metastasis, such as Krukenberg tumor, peritoneal metastasis, or lymph node metastasis).