JPH03240498A - Antibody and immune analysis using same antibody - Google Patents
Antibody and immune analysis using same antibodyInfo
- Publication number
- JPH03240498A JPH03240498A JP2033977A JP3397790A JPH03240498A JP H03240498 A JPH03240498 A JP H03240498A JP 2033977 A JP2033977 A JP 2033977A JP 3397790 A JP3397790 A JP 3397790A JP H03240498 A JPH03240498 A JP H03240498A
- Authority
- JP
- Japan
- Prior art keywords
- antibody
- peptide
- monoclonal antibody
- erbb
- mouse
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 28
- 101710100968 Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 claims abstract description 18
- 102000036639 antigens Human genes 0.000 claims abstract description 17
- 108091007433 antigens Proteins 0.000 claims abstract description 17
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 claims abstract description 16
- 239000000427 antigen Substances 0.000 claims abstract description 15
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- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 5
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- 238000003018 immunoassay Methods 0.000 claims description 5
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- 238000002360 preparation method Methods 0.000 abstract description 4
- 206010035226 Plasma cell myeloma Diseases 0.000 abstract description 3
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- 229940125396 insulin Drugs 0.000 abstract description 2
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- 102000004196 processed proteins & peptides Human genes 0.000 description 9
- 239000000032 diagnostic agent Substances 0.000 description 6
- 229940039227 diagnostic agent Drugs 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 208000009956 adenocarcinoma Diseases 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 5
- 206010006187 Breast cancer Diseases 0.000 description 4
- 208000026310 Breast neoplasm Diseases 0.000 description 4
- 241000699670 Mus sp. Species 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 102000014914 Carrier Proteins Human genes 0.000 description 3
- 108010078791 Carrier Proteins Proteins 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 3
- 230000007910 cell fusion Effects 0.000 description 3
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- 108700020796 Oncogene Proteins 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
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- 108010045069 keyhole-limpet hemocyanin Proteins 0.000 description 2
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- 229920001223 polyethylene glycol Polymers 0.000 description 2
- XOJVVFBFDXDTEG-UHFFFAOYSA-N pristane Chemical compound CC(C)CCCC(C)CCCC(C)CCCC(C)C XOJVVFBFDXDTEG-UHFFFAOYSA-N 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
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- 206010003445 Ascites Diseases 0.000 description 1
- -1 CVGEGL Proteins 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
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- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
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Abstract
Description
【発明の詳細な説明】
「産業上の利用分野］
本発明は、腺癌の癌化と密接に関係して発現すると予想
されるヒト癌遺伝子ｃ−ｅｒｂＢ−２産物に特異的なモ
ノクローナル抗体およびそれを用いた免疫分析方法に関
するものであり、当該抗体を放射性同位元素、蛍光色素
、酵素などで標識し、腺癌の病巣部位や転移の診断を行
なう診断薬として用いることができる。特に本抗体の特
徴を生かし画像診断薬として有用である。さらに、この
抗体は、イムノトキシンとしても応用されることが期待
される。Detailed Description of the Invention "Industrial Application Field" The present invention provides a monoclonal antibody and a monoclonal antibody specific for the human oncogene c-erbB-2 product, which is predicted to be expressed in close association with the carcinogenesis of adenocarcinoma. This antibody relates to an immunoassay method using the antibody, and the antibody can be labeled with a radioactive isotope, a fluorescent dye, an enzyme, etc., and used as a diagnostic agent for diagnosing the focal site and metastasis of adenocarcinoma.In particular, this antibody Taking advantage of its characteristics, this antibody is useful as an imaging diagnostic agent.Furthermore, this antibody is expected to be applied as an immunotoxin.
［従来技術、発明が解決しようとする課題］腺癌の診断
法としては、超音波によるものや、金コロイドあるいは
テクネシウム放射性同位元素の血中投与によるもの等が
あるが、前者は癌腫瘍とリンパ部の腫れ等の区別ができ
ず、後者は単に血管分布を調べているため癌腫瘍と血管
部位との区別ができない。すなわち、これらの方法は癌
腫瘍に対する特異性の面で欠点がある。一方、癌組織を
特異的に識別し得るものとして抗体が上げられ、腺癌の
画像診断薬としては、治験中ではあるが、ＣＥＡを抗原
としたものがある。現在、本発明に類似するｃ−ｅｒｂ
Ｂ−２産物に対する抗体作製の報告が数例あるが、その
ほとんどは細胞内のリン酸化部位と反応するものであり
、生体での癌組織の認識はできない。下記本発明の抗体
は、細胞外の部位と反応し、さらに癌組織の免疫染色が
可能であり、画像診断薬として利用できる抗体としは他
に例がない。[Prior Art and Problems to be Solved by the Invention] Methods for diagnosing adenocarcinoma include ultrasound, and administration of colloidal gold or technetium radioisotope into the blood. The latter cannot distinguish between cancerous tumors and vascular sites because it merely examines blood vessel distribution. That is, these methods have shortcomings in terms of specificity for cancer tumors. On the other hand, antibodies are cited as something that can specifically identify cancerous tissue, and there is an imaging diagnostic agent for adenocarcinoma that uses CEA as an antigen, although it is currently under clinical trials. Currently, c-erb similar to the present invention
Although there are several reports on the production of antibodies against B-2 products, most of them react with intracellular phosphorylation sites and cannot recognize cancer tissues in living organisms. The antibody of the present invention described below reacts with extracellular sites and is also capable of immunostaining cancer tissues, and is unique as an antibody that can be used as an imaging diagnostic agent.
［課題を解決するための手段］
癌組織を効果的に識別し得る抗体を得るために、いろい
ろな腺癌組織において増幅されている癌遺伝子ｃ−ｅｒ
ｂＢ−２に着目した。ｃ−ｅｒｂＢ−２は、特に乳癌組
織あるいは胃癌組織において増幅されていることが知ら
れている。従って、ｃ−ｅｒｂＢ−２の発現とこれらの
癌の癌化には、密接な関係があるものと考えられる。ま
た、ｃ−ｅｒｂＢ−２の産物は、分子量１８５ｋＤａの
膜貫通性の蛋白質であることが知られているので、細胞
外部に貫通したｃ−ｅｒｂＢ−２の産物の部位を認識す
る抗体を見出すことにより、非常に有益な抗体が得られ
ると予想される。[Means for solving the problem] In order to obtain antibodies that can effectively identify cancer tissues, the oncogene c-er, which is amplified in various adenocarcinoma tissues, is
We focused on bB-2. It is known that c-erbB-2 is particularly amplified in breast cancer tissue or gastric cancer tissue. Therefore, it is thought that there is a close relationship between the expression of c-erbB-2 and the transformation of these cancers. Furthermore, since the product of c-erbB-2 is known to be a transmembrane protein with a molecular weight of 185 kDa, it is important to find an antibody that recognizes the site of the product of c-erbB-2 that penetrates to the outside of the cell. It is expected that very useful antibodies will be obtained.
そこで、ｃ−ｅｒｂＢ−２産物の蛋白質アミノ酸配列の
中で、細胞外の部分に相当する親水性部位のペプチドを
いくつか合成し、それを抗原として得た抗体を評価した
。Therefore, in the protein amino acid sequence of the c-erbB-2 product, several peptides with hydrophilic sites corresponding to the extracellular portion were synthesized, and antibodies obtained using the peptides as antigens were evaluated.
即ち、後述第１表記載のアミノ酸残基数的１０〜２０の
親水性ペプチドを合成し、それを抗原として通例の方法
によりポリクローナル抗体を得、それを用いて癌細胞の
免疫分析を行った。その結果、いずれも反応性に差異の
あるものの癌細胞を特異的に認識しつる抗体であること
がわかった。That is, a hydrophilic peptide having 10 to 20 amino acid residues listed in Table 1 below was synthesized, a polyclonal antibody was obtained using the peptide as an antigen by a conventional method, and immunoassay of cancer cells was performed using the peptide. As a result, they were all found to be antibodies that specifically recognized cancer cells, although their reactivity differed.
そこで本発明者は、さらに抗体の特異性を高めるために
上記抗原を用いてモノクローナル抗体を作威し、それを
評価した。その結果、特異性が極めて高いモノクローナ
ル抗体を得ることができた。Therefore, in order to further enhance the specificity of the antibody, the present inventor created a monoclonal antibody using the above antigen and evaluated it. As a result, a monoclonal antibody with extremely high specificity could be obtained.
本発明は、上記モノクローナル抗体およびそれを使用し
た免疫分析方法に係る発明である。The present invention relates to the above monoclonal antibody and an immunoassay method using the same.
本発明において、抗原のペプチドとしては、後述第１表
５に記載のペプチドが最も好ましい。しかし、これに限
られるものではなく、第１表１．および２．に記載のペ
プチドも好ましいペプチドである。また、抗原は通常キ
ャリアー蛋白質に結合させて抗体製造に用いられる。In the present invention, as the antigen peptide, the peptides listed in Table 1 below are most preferred. However, it is not limited to this, and Table 1 1. and 2. The peptides described in are also preferred peptides. Further, antigens are usually bound to carrier proteins and used for antibody production.
なお、本発明におけるモノクローナル抗体は後述実施例
に示すように、公知のモノクローナル抗体製造法で製造
することができるものである。The monoclonal antibody in the present invention can be produced by a known monoclonal antibody production method, as shown in Examples below.
参考例
ｃ −ｅ ｒ Ｉ）Ｂ −？、癌遺伝子によってコード
された産物の親水性部位をＣｈｏｕ−Ｆａｓｍａｎの方
法を用いて検索し、それらのいくつかについて、その合
成ペプチドを作成し、キャリアー蛋白質に結合し、それ
を抗原としてマウスを免疫した。得られた抗血清につい
て、ｃ−ｅｒｂＢ−２の発現が認識されている乳癌細胞
株Ｓ Ｋ −Ｂ Ｒ−Ｉｌｌに対する反応性を、細胞を
固相化したＥＬＪＳＡ法で調べ、第１表の結果を得た。Reference example c -e r I) B -? We searched for hydrophilic sites of products encoded by oncogenes using Chou-Fasman's method, created synthetic peptides for some of them, bound them to carrier proteins, and immunized mice with them as antigens. did. The reactivity of the obtained antiserum against the breast cancer cell line SK-BR-Ill, which is known to express c-erbB-2, was examined using the ELJSA method using immobilized cells, and the results are shown in Table 1. I got it.
この結果から、第１表の５の成分ペプチドが最も有効で
あることが見出された。From this result, it was found that component peptide 5 in Table 1 was the most effective.
第１表 親水性部位のペプチドと反応性実施例
ｃ−ｅｒｂＢ−２癌遺伝子によってコードされた産物の
Ｎ端４９５から５０９までの残基のペプチド（上記第１
表５の合成ペプチド）を抗原とし、それにより免疫した
マウス牌細胞と、マウス骨髄腫細胞（５Ｐ２１０）を細
胞融合させ、ハイブリドーマを作製することにより得ら
れる。作製手順の概要は以下である。Table 1 Reactivity with peptides in the hydrophilic region Example Peptide of residues 495 to 509 at the N terminus of the product encoded by the c-erbB-2 oncogene (the first
Synthetic peptides shown in Table 5) are used as antigens, and mouse tile cells immunized with the antigens are fused with mouse myeloma cells (5P210) to produce hybridomas. The outline of the manufacturing procedure is as follows.
抗原ペプチドは、アミノ酸配列がＩ（ＴＡＮＲＰＥＤＥ
ＣＶＧＥＧＬである合成ペプチドと、キャリヤー蛋白質
として、キーホールリンペットヘモシアニン（ＫＬＨ）
をグルタルアルデヒドを用いて架橋させて作製した。作
製の方法ば、Ｇ、 Ｗａｉｔｅｒ等の方法（Ｐｒｏｃ、
Ｎａｔｌ、 Ａｃａｄ、 Ｓｃｉ、 、 ７７、６１
９７．１９８０）に準じた。次に、免疫は、上記を抗原
としＢａ１ｂ／Ｃマウスを用いた。また、細胞融合は、
ＢｅｈｚａｄｉａｎＭ、Ａ、等の方法（Ｃｅｌｌ ５ｔ
ｒｕｃｔ、Ｆｕｎｃｔ、、１０．２１９１９８５）に準
じて行なった。スクリーニングはＥＬＩＳＡ法を用いた
。The antigenic peptide has an amino acid sequence of I (TANRPEDE
A synthetic peptide, CVGEGL, and keyhole limpet hemocyanin (KLH) as a carrier protein.
was prepared by crosslinking using glutaraldehyde. The method of preparation is the method of G. Waiter et al. (Proc,
Natl, Acad, Sci, 77, 61
97.1980). Next, Ba1b/C mice were immunized using the above antigen as an antigen. In addition, cell fusion
The method of Behzadian M, A, et al. (Cell 5t
ruct, Funct, 10.2191985). Screening used the ELISA method.
得られたモノクローナル抗体はヒトｃ−ｅｒｂＢ−２産
物を認識する。なお、このモノクローナル抗体が、ｃ−
ｅｒｂＢ−２産物に対するものであるか否かは、ｃ−ｅ
ｒｂＢ−２の発現が確認されている乳癌細胞株Ｓ Ｋ
−Ｂ Ｒ−ＩＩＩの蛋白質を３’ Ｓ −ｍ ｅ ｔ
ｈ ｉ ｏ ｎ ｉ ｎ ｅで放射標識したものを可溶
化し免疫沈澱を行ない、ＳＤＳ電気泳動法で展開後、そ
のオートラジオグラフィーを分析することにより、確認
された。The resulting monoclonal antibody recognizes the human c-erbB-2 product. Note that this monoclonal antibody is c-
Whether it is for the erbB-2 product or not, c-e
Breast cancer cell line SK with confirmed expression of rbB-2
-B R-III protein to 3' S -met
This was confirmed by solubilizing radiolabeled hionine, performing immunoprecipitation, developing it by SDS electrophoresis, and analyzing its autoradiography.
■、モノクローナル抗体の製造
（１）抗原蛋白質の調製
前記ペプチドの合成は、自動ペプチド合成装置による固
相法で行なった。さらに、］、ｍｇＫｌ、１１と３ｍｇ
ペプチドを０．］、Ｍリン酸緩衝液（ｐ、 Ｈ７，２）
中で、２．５％グルタルアルデヒドで架橋反応を行なっ
た。さらに、フリーのグルタルアルデヒドを除くために
、０．０１Ｍリン酸緩衝液（ｐＨ７，４）＋０．１５Ｍ
ＮａＣ１中で透析を行なった。(2) Production of monoclonal antibodies (1) Preparation of antigen protein The synthesis of the peptides described above was carried out by a solid phase method using an automatic peptide synthesizer. Furthermore, ], mgKl, 11 and 3 mg
0.0 peptide. ], M phosphate buffer (p, H7,2)
A crosslinking reaction was carried out using 2.5% glutaraldehyde. Furthermore, to remove free glutaraldehyde, 0.01M phosphate buffer (pH 7.4) + 0.15M
Dialysis was performed in NaCl.
（２）免疫
１００μｇ抗原蛋白質とフロイント、不完全アジュバン
トを、Ｂａ１ｂ／ｃマウスの皮下に１４日間隔で３回投
与した。ペプチドに対する抗血清が得られているか否か
は、ペプチドを９６穴プ１ノートに固定しＥＬＩＳＡ法
で調べた。この時、抗マウスＩｇＧに酵素ＰＯＸを接合
したものを用いた。基質どしてはＯＰＤである。(2) Immunization 100 μg of antigen protein and Freund's incomplete adjuvant were subcutaneously administered to Ba1b/c mice three times at 14-day intervals. Whether antiserum against the peptide was obtained was determined by immobilizing the peptide in a 96-well plate and using ELISA. At this time, anti-mouse IgG conjugated with the enzyme POX was used. The substrate is OPD.
（３）細胞融合
免疫されたＢａ１ｂ／ｃマウスの牌蔵を摘出し、ＲＰＭ
Ｉ ］、６４０培地中で牌細胞を押し出す。マウス骨髄
腫細胞５Ｐ２１０を牌細胞数の１１５〜１／ｌＯ量加え
、ＲＰＭＩを遠心で除いた後、ポリエチレングリコール
（分子量４０００、シグマ）で細胞融合を行なった。ポ
リエチレングリコールの排除、融合の確実性を得るため
に、直ちに遠心を行ない、ＨＡＴ培地にて培養した。約
−週間後血清培地にて限界希釈法にてクローニングを行
なった。スクリーニングはＥＬＩＳＡ法にて行なった。(3) Excise the tile of Ba1b/c mouse immunized with cell fusion, and RPM
I], extrude tile cells in 640 medium. Mouse myeloma cells 5P210 were added in an amount of 115 to 1/lO of the number of tile cells, and after removing RPMI by centrifugation, cell fusion was performed with polyethylene glycol (molecular weight 4000, Sigma). In order to eliminate polyethylene glycol and ensure fusion, the cells were immediately centrifuged and cultured in HAT medium. Approximately one week later, cloning was carried out using the limiting dilution method in a serum medium. Screening was performed by ELISA method.
（４）モノクローナル抗体の調製
得られたハイブリドーマを、１５％ＦＣ３、グルタミン
酸、インシュリンを含んだＲＰＭＩ １６４０培地で増
殖し、Ｘ線処理、プリスタン投与したＢａ１ｂ／ｃマウ
スの腹腔内に細胞（１０７／匹）を投与する。約３週間
後、腹水を取り遠心分離を行ない、上清からＤＥＡＥセ
ルロースカラムを用いて抗体を精製した。(4) Preparation of monoclonal antibodies The obtained hybridomas were grown in RPMI 1640 medium containing 15% FC3, glutamic acid, and insulin, treated with X-rays, and intraperitoneally injected into Ba1b/c mice (107 cells/mouse) administered with pristane. ). After about 3 weeks, ascites was collected and centrifuged, and antibodies were purified from the supernatant using a DEAE cellulose column.
ＩＩ 、 ｃ−ｅｒｂＢ−２産物に対するモノクローナ
ル抗体の性質
（１）抗体のサブクラスの固定
０ｕｃｈｔｅｒ］、ｏｎｙ法に準じて行なった。抗マウ
スＩｇＭ又は抗マウスＩｇＧと、得られたモノクローナ
ル抗体を１％寒天中で反応させ、沈降線を生ずるか否か
で調べた結果、モノクローナル抗体はＩｇＭと確認され
た。II. Properties of monoclonal antibodies against c-erbB-2 products (1) Immobilization of antibody subclasses It was carried out according to the ony method. The obtained monoclonal antibody was reacted with anti-mouse IgM or anti-mouse IgG in 1% agar, and the monoclonal antibody was confirmed to be IgM by examining whether a sedimentation line was produced.
（２）分子量
セファクリルＳ−３００５ｕｐｅｒｆｉｎｅを用いてカ
ラムクロマトグラフィーで調べた。(2) Molecular weight was investigated by column chromatography using Sephacryl S-3005 upperfine.
（３）抗体の免疫特性
”Ｓ−ｍｅｔｈｉｏｎｉｎｅで蛋白質を放射標識したＳ
Ｋ −Ｂ Ｒ−ＩＩＩ細胞を可溶化し、それにモノク
ローナル抗体を加え反応させ、遠心による沈澱物の数回
の洗浄後、ＳＤＳ電気泳動（７，５％）で展開し、その
ゲルのオートラジオグラフィーを取ったところ、１８５
ｋＤａ例近に一本のバンドが観察された。この分子量は
ヒ１〜ｃ−ｅｒｂＢ−２産物と同じであり、この抗体が
ｃ−ｅｒｂＢ−２産物を認識していることかわかる。(3) Immunological properties of antibodies: S-methionine radiolabeled protein
K-B R-III cells were solubilized, a monoclonal antibody was added thereto, and the precipitate was washed several times by centrifugation, developed by SDS electrophoresis (7.5%), and the gel was autoradiographed. However, 185
A single band was observed near kDa. This molecular weight is the same as that of the human c-erbB-2 product, indicating that this antibody recognizes the c-erbB-2 product.
乳癌組織を採取後、直ちにＯ，Ｃ，Ｔ、 ｃｏｍｐｏｕ
ｎｄに包埋凍結し、新鮮凍結切片を作成し、ＡＢＣ法に
て免疫染色を行なった。その結果癌包巣の細胞は良く染
色されたが、それ以外の部分では染色性が認められなか
った。Immediately after collecting breast cancer tissue, O, C, T, compou
The cells were embedded and frozen, fresh frozen sections were prepared, and immunostaining was performed using the ABC method. As a result, cells in the cancer fodder were well stained, but no staining was observed in other areas.
ここで、免疫原である合成ペプチドで抗体と競合させる
と、癌組織はほとんど染色されなかった。また、１５残
基からなる任意の合成ペプチドを加えても、染色性には
影響なく、この抗体の代りにノーマル抗体で処理すると
、全く染色されなかった。Here, when a synthetic peptide, which is an immunogen, was used to compete with the antibody, the cancer tissue was hardly stained. Furthermore, addition of an arbitrary synthetic peptide consisting of 15 residues had no effect on the staining properties, and when treated with a normal antibody instead of this antibody, no staining was observed at all.
以上のことは、このモノクローナル抗体が画像診断薬と
して利用できることを示している。The above indicates that this monoclonal antibody can be used as an imaging diagnostic agent.
［発明の効果］
本発明のモノクローナル抗体は、ｃ−ｅｒｂＢ−２産物
に対する特異性が高く、しかもｃ−ｅｒｂＢ−２産物の
細胞外表面に存在する部位を認識する抗体である。従っ
て、本発明の抗体は生体組織中のｃ−ｅｒｂＢ−２産物
を発現した癌細胞を他の細胞から区別して認識すること
ができ、免疫染色による画像診断薬どして優れた特性を
有する。[Effects of the Invention] The monoclonal antibody of the present invention has high specificity for the c-erbB-2 product, and is an antibody that recognizes a site present on the extracellular surface of the c-erbB-2 product. Therefore, the antibody of the present invention can distinguish and recognize cancer cells expressing c-erbB-2 products from other cells in living tissues, and has excellent properties as an imaging diagnostic agent using immunostaining.
１ ２1 2
Claims (1)
胞外親水部位の部分ペプチドを抗原として得られたモノ
クローナル抗体。 ２、抗原が下記アミノ酸配列のペプチドである、請求項
第１項記載のモノクローナル抗 体。 Ｈｉｓ−Ｔｈｒ−Ａｌａ−Ａｓｎ−Ａｒｇ−Ｐｒｏ−Ｇ
ｌｕ−Ａｓｐ−Ｇｌｕ−Ｃｙｓ−Ｖａｌ−Ｇｌｙ−Ｇｌ
ｕ−Ｇｌｙ−Ｌｅｕ ３、請求項第１項記載のモノクローナル抗体を使用して
免疫分析を行うことを特徴とする免疫分析方法。 ４、癌細胞を識別する、請求項第３項記載の方法。[Scope of Claims] 1. A monoclonal antibody obtained using a partial peptide of the extracellular hydrophilic site in the protein amino acid sequence of the c-erbB-2 product as an antigen. 2. The monoclonal antibody according to claim 1, wherein the antigen is a peptide having the following amino acid sequence. His-Thr-Ala-Asn-Arg-Pro-G
lu-Asp-Glu-Cys-Val-Gly-Gl
An immunoassay method comprising performing an immunoassay using u-Gly-Leu 3, the monoclonal antibody according to claim 1. 4. The method according to claim 3, wherein cancer cells are identified.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2033977A JPH03240498A (en) | 1990-02-16 | 1990-02-16 | Antibody and immune analysis using same antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2033977A JPH03240498A (en) | 1990-02-16 | 1990-02-16 | Antibody and immune analysis using same antibody |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH03240498A true JPH03240498A (en) | 1991-10-25 |
Family
ID=12401550
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2033977A Pending JPH03240498A (en) | 1990-02-16 | 1990-02-16 | Antibody and immune analysis using same antibody |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH03240498A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5681729A (en) * | 1991-10-30 | 1997-10-28 | Idemitsu Kosan Company Limited | Methods for producing human lymphocytes and human monoclonal antibodies, and human monoclonal antibodies produced thereby |
| US6627196B1 (en) | 1999-08-27 | 2003-09-30 | Genentech, Inc. | Dosages for treatment with anti-ErbB2 antibodies |
| US7041292B1 (en) | 1999-06-25 | 2006-05-09 | Genentech, Inc. | Treating prostate cancer with anti-ErbB2 antibodies |
| US7371376B1 (en) | 1996-10-18 | 2008-05-13 | Genentech, Inc. | Anti-ErbB2 antibodies |
| US10501521B2 (en) | 2012-12-21 | 2019-12-10 | Hoffmann-La Roche Inc. | Disulfide-linked multivalent MHC class I comprising multi-function proteins |
-
1990
- 1990-02-16 JP JP2033977A patent/JPH03240498A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5681729A (en) * | 1991-10-30 | 1997-10-28 | Idemitsu Kosan Company Limited | Methods for producing human lymphocytes and human monoclonal antibodies, and human monoclonal antibodies produced thereby |
| US5869268A (en) * | 1991-10-30 | 1999-02-09 | Idemitsu Kosan Company Limited | Methods for producing human lymphocytes and human monoclonal antibodies, and human monoclonal antibodies produced thereby |
| US7371376B1 (en) | 1996-10-18 | 2008-05-13 | Genentech, Inc. | Anti-ErbB2 antibodies |
| US7041292B1 (en) | 1999-06-25 | 2006-05-09 | Genentech, Inc. | Treating prostate cancer with anti-ErbB2 antibodies |
| US6627196B1 (en) | 1999-08-27 | 2003-09-30 | Genentech, Inc. | Dosages for treatment with anti-ErbB2 antibodies |
| US7371379B2 (en) | 1999-08-27 | 2008-05-13 | Genentech, Inc. | Dosages for treatment with anti-ErbB2 antibodies |
| US10160811B2 (en) | 1999-08-27 | 2018-12-25 | Genentech, Inc. | Treatment with anti-ErbB2 antibodies |
| US10280228B2 (en) | 1999-08-27 | 2019-05-07 | Genentech, Inc. | Treatment with anti-ErbB2 antibodies |
| US10501521B2 (en) | 2012-12-21 | 2019-12-10 | Hoffmann-La Roche Inc. | Disulfide-linked multivalent MHC class I comprising multi-function proteins |
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