CN1683409A - Single strand double specificity anti-body resisting human B cell leucoma - Google Patents
Single strand double specificity anti-body resisting human B cell leucoma Download PDFInfo
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- CN1683409A CN1683409A CN 200510051233 CN200510051233A CN1683409A CN 1683409 A CN1683409 A CN 1683409A CN 200510051233 CN200510051233 CN 200510051233 CN 200510051233 A CN200510051233 A CN 200510051233A CN 1683409 A CN1683409 A CN 1683409A
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Abstract
The present invention discloses a kind of single strand double specificity antibody resisting human B cell leucoma. Three interchain connecting peptides (Gly4Ser)3 are used to connect the light strand variable area VLCD20 and heavy strand variable area VHCD20 of human B cell leucoma CD20 resisting antibody as well as the light strand variable area VLCD3 and heavy strand variable area VHCD3 of human CD3 resisting antibody to form VLCD3-(Gly4Ser)3-VHCD3-(Gly4Ser)3-VHCD20-(Gly4Ser)3-VLCD20 single strand antibody. The single strand antibody is one single strand double specificity antibody capable of combining with CD3 and CD20 specifically, mediating human peripheral blood lymphocyte to kill human B cell leucoma and resisting CD3/CD20.
Description
Technical field
The present invention relates to a kind of genetic engineering antibody, particularly relate to the single chain bispecific antibody of target killing B lymphoma cell with anti-tumor activity.
Background technology
(single chain bispecific antibody ScBsAb) is a kind of non-natural genetic engineering antibody form to single chain bispecific antibody, and two has different specificitys by the antigen-binding site that connection peptides links to each other.Therefore, though ScBsAb is two valencys on chemical structure, with regard to the function of its conjugated antigen, be monovalent.The specific antibody of tumor associated antigen and the surface of tumor-killing cell are triggered the ScBsAb of the specific antibody of molecule by the connection peptides formation, can efficiently intravital immune effector cell be raised around tumour cell, the activate immunity effector cell is killing tumor cell specifically, thereby reaches the purpose of treatment tumour.
CD20 is a kind of nonglycosylated III type transmembrane protein, the conservative property of tool height.CD20 only expresses in pre-B lymphocyte, immature B lymphocyte, ripe bone-marrow-derived lymphocyte and activated bone-marrow-derived lymphocyte, and does not all have expression in plasmocyte, lymph multipotential stem cell and other tissue.Surpass 95% B cell lymphoma expression CD20, and do not have tangible CD20 endocytosis or obscission, these features to make CD20 become the desirable target antigen of treatment B cell lymphoma.Retuximab (in December, 1997) and Ibritumomal (in February, 2002) that drugs approved by FDA is used for the treatment of B cell lymphoma promptly are respectively at the chimeric IgG of people-mouse of CD20
1Type antibody and isotope-labeled mouse source property IgG
1The type monoclonal antibody.
Effector cell's toxicity T cell is the strongest " killer " in the human immunity safeguard system.The major function of T cell is identification and promptly destroys those by virus infection or suffer the cell or tissue of vicious transformation.The activation of T cell is the basic prerequisite of its performance cell killing effect.The activation of T cell is not simple single step reaction, but is subjected to the control of a series of cell surface protein, guarantees that with this T cell only acts on specific target.These surface proteins comprise TXi Baoshouti, CD3 signal mixture, costimulatory molecules and the adhesion molecule that can discern target cell surface antigen and MHC I quasi-molecule mixture specifically.Under their effect, form an immunity between T cell and the target cell and connect, static T cell is activated, thus the effect that performance kills and wounds target cell.In addition, tumour cell also may tackle the lethal effect of T cell by " immune evasion " strategy, as reducing the expression of tumor cell surface MHC I quasi-molecule, makes it and can not form mixture with surface antigen, and then can not be discerned by TXi Baoshouti.The complexity regulation and control and the escape mechanism of tumor cells of t cell activation process have limited giving full play to of T cell killing tumour cell potential greatly.
BiTEs (bispecific T cell engagers) is to be effector cell's single chain bispecific antibody with the T cell.The difference of BiTEs and other bi-specific antibody maximum is its structure, and it adopts the single-chain antibody technology that the antigen-binding site of two different antibodies is linked together by connection peptides, and its basic form is VL
1-linker-VH
1-linker-VH
2-linker-VL
2This structure is given between two brachium conjunctivums of BiTEs has better snappiness, can be simultaneously and the antigen molecule combination on T cell and target cell surface, and the toxicity that forms effective activated T cell between T cell and target cell connects, and mediates the cracking of target cell.
BiTEs is as a kind of gene engineering single chain bi-specific antibody, in the tumor-killing effect of fully having excavated the cytotoxic T cell that enriches in the human body, overcome the defective that the whole antibody molecular weight is big, penetrativity is low, good potential applicability in clinical practice has been arranged in the biotherapy of tumour.
The single chain bispecific antibody that the anti-CD20 single chain bispecific antibody of anti-CD3/ promptly is a kind of BiTEs form, the specificity combination can take place with specific antigens molecule CD3, the CD20 on T cell, B lymphoma cell surface in it, for a physical connection is provided between T cell and the B lymphoma cell, activate immobilized T cell killing tumour cell effectively, play the effect of treatment B cell lymphoma.
At present, the single chain bispecific antibody relevant for the BiTEs form all has report both at home and abroad.Wherein, the development of German Micromet company has got permission to enter the clinical experimental study that is used for the treatment of Fei Hejiejinshi B cell lymphoma and lymphocytic leukemia at the antigenic BiTEs of B cell surface CD19 (MT103).At the antigenic BiTEs of CD20, the anti-CD20 single chain bispecific antibody of promptly anti-CD3/ there is no report both at home and abroad.
Summary of the invention
The invention provides a kind of is single chain bispecific antibody target, that have target killing B cell lymphoma function (the anti-CD20 single chain bispecific antibody of anti-CD3/) with CD20.
The present invention includes following main contents:
The anti-CD20 single chain bispecific antibody of anti-CD3/ of the present invention is made of anti-human B cell lymphoma CD20 single-chain antibody and anti-people CD3 single-chain antibody, specifically will resist the variable region of light chain (VL of human B cell lymphoma CD20 antibody by 3 interlinkers
CD20) and variable region of heavy chain (VH
CD20) and the variable region of light chain (VL of anti-people CD3 antibody
CD3) and variable region of heavy chain (VH
CD3) be formed by connecting.It can effectively activate the lymphadenomatous effect of immobilized T lymphocyte performance specific killing human B cell.
The anti-CD20 single chain bispecific antibody of anti-CD3/ of the present invention adopts (Gly
4Ser)
3Be connection peptides, its type of attachment is VL
CD3-(Gly
4Ser)
3-VH
CD3-(Gly
4Ser)
3-VH
CD20-(Gly
4Ser)
3-VL
CD20
In the anti-CD20 single chain bispecific antibody of anti-CD3/ of the present invention, anti-human B cell lymphoma CD20 single-chain antibody has the weight chain variable region amino acid sequence shown in sequence table SEQ ID No.1.
In the anti-CD20 single chain bispecific antibody of anti-CD3/ of the present invention, anti-human B cell lymphoma CD20 single-chain antibody has the light chain variable region amino acid sequence shown in sequence table SEQ ID No.2:
In the anti-CD20 single chain bispecific antibody of anti-CD3/ of the present invention, anti-people CD3 single-chain antibody has the weight chain variable region amino acid sequence shown in sequence table SEQ ID No.3:
In the anti-CD20 single chain bispecific antibody of anti-CD3/ of the present invention, anti-people CD3 single-chain antibody has the light chain variable region amino acid sequence shown in sequence table SEQ ID No.4:
The present invention compares following advantage with existing at the antigenic other forms of antibody of CD20: have better snappiness between two antigen brachium conjunctivums in the anti-CD20 single chain bispecific antibody of 1. anti-CD3/, this snappiness helps promoting the connection of CD3 complex body and tumour target and form toxicity at last between T cell and target cell connecting, thereby activates immobilized T cell killing tumour cell effectively.2. the anti-CD20 single chain bispecific antibody of anti-CD3/ molecular weight is less, stronger to the penetrativity of tumor tissues.3. the structure of the anti-CD20 single chain bispecific antibody of CD3/ is simpler, is convenient to adopt the biotechnology means to prepare in a large number.
The invention will be further described below in conjunction with drawings and Examples.
Description of drawings
Fig. 1 is the physical map of expression plasmid pcDNA5/FRT-amiCD3/antiCD20.
Fig. 2 is for adopting viable cell indirect immunofluorescence detection method observed Ramous cell (2A) and Jurkat cell (2B) under fluorescent microscope.
Fig. 3 is under the single dual specific chain antibody mediation of the anti-CD20 of anti-CD3/, by Jurkat cell and the plastidogenetic rosettes of Ramous.
Fig. 4 is that human peripheral lymphocyte under the mediation of the anti-CD20 single chain bispecific antibody of anti-CD3/ is to the lethal effect of people B lymphoma cell.
Embodiment
One. the design of the anti-CD20 single chain bispecific antibody of anti-CD3/ nucleotide sequence and synthetic
According to VL
CD3, VH
CD3, VL
CD20, VH
CD20Aminoacid sequence and VL
CD3-(Gly
4Ser)
3-VH
CD3-(Gly
4Ser)
3-VH
CD20-(Gly
4Ser)
3-VL
CD20Type of attachment, determine the nucleotide sequence of the anti-CD20 single chain bispecific antibody of anti-CD3/ with reference to mammalian cell codon commonly used.For the ease of proteic purifying, introduce histidine-tagged, NotI restriction enzyme site at 3 ' end of sequence, and histidine-tagged before adding zymoplasm restriction enzyme site so that finally remove histidine-tagged; 5 ' end in sequence adds Nhe I restriction enzyme site, KOZAK sequence and leader sequence.
The oligonucleotide fragment of the synthetic a plurality of about 60bp of design, every two adjacent fragments base complementrity of 20bp of having an appointment, having obtained total length by overlapping extension PCR (SOE PCR) is the anti-CD20 single chain bispecific antibody of the anti-CD3/ complete genome sequence of 1640bp.The full gene the primer that increases is: upstream primer: 5 '-CTAGCTAGCG CCGCCAGCC-3 '.Downstream primer: 5 '-GTTTTCCTTT TGCGGCCGCC TA-3 '.
The anti-CD20 single chain bispecific antibody of anti-CD3/ complete genome sequence synoptic diagram is as follows:
-----KOZAK sequence-leader sequence-VL
CD3-(G
4S)
3-VH
CD3-(G
4S)
3-VH
CD20-(G
4S)
3-VL
CD20-blood coagulation restriction enzyme site-(His)
6-----.
Two. the structure of the anti-CD20 single chain bispecific antibody of anti-CD3/ expression vector
Behind the full gene usefulness Nhe I and Not I double digestion with the anti-CD20 single chain bispecific antibody of anti-CD3/, be connected to through Nhe I and Not I enzyme with the T4 ligase enzyme and cut among the eucaryon fixed point expression vector pcDNA5/FRT (available from American I nvitrogen company) of processing, obtain the expression vector pcDNA5/FRT-antiCD3/antiCD20 (Fig. 1) of the anti-CD20 single chain bispecific antibody of anti-CD3/.With pcDNA5/FRT-antiCD3/antiCD20 transformed into escherichia coli DH5 α, the picking positive colony carries out enzyme through bacterium colony and cuts evaluation and sequencing analysis.
Three. the expression of the anti-CD20 single chain bispecific antibody of anti-CD3/ in Chinese hamster ovary celI
PcDNA5/FRT-antiCD3/antiCD20 expression plasmid that the employing liposome method will check order correct and pOG44 plasmid (available from American I nvitrogen company) the cotransfection Flp-In of express recombinant enzyme Flp
TMCHO transfectional cell (available from American I nvitrogen company), and be Hgromycm B (available from the American I nvitrogen company) screening positive clone of 500 μ g/ml with concentration.Adopt direct competitive ELISA to detect the expression of the anti-CD20 single chain bispecific antibody of anti-CD3/.
Four. the purifying of the anti-CD20 single chain bispecific antibody of anti-CD3/
The positive colony cell that in rolling bottle and blender jar, screens with the serum free medium amplification culture, the collecting cell culture supernatant, adding final concentration is the imidazoles of 5mM and the CaCl of 1mM
2, by Ni-NTA chromatography column (GE Helthcare company), earlier with the PBS wash-out foreign protein that contains 0.5M NaCl, the PBS that is respectively 20mM to 500mM with imidazole concentration carries out the continuous gradient wash-out to target protein again with the flow velocity of 1ml/min.
The biological activity of the anti-CD20 single chain bispecific antibody of embodiment 2. anti-CD3/ is identified
One. the antigen-binding activity of the anti-CD20 single chain bispecific antibody of anti-CD3/
With the anti-CD20 single chain bispecific antibody of the anti-CD3/ of certain density purifying respectively with 1 * 10
6Ramous cell or Jurkat cell (all available from Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences's cell centre) are hatched 1h for 4 ℃ jointly, 4 ℃ of centrifugal 3min of 3000r/min, and PBS washes 3 times.Anti-His (the C-term)-FITC antibody (available from American I nvitrogen company) that adds 50 μ l dilution in 1: 500,4 ℃ of lucifuge effect 1h, 4 ℃ of centrifugal 3min of 3000r/min, PBS washes 4 times, and fluorescent microscope is observed down and is taken a picture.Under fluorescent microscope, Ramous cell and Jurkat cell all manifest tangible green fluorescence (Fig. 2), and mainly are distributed in around the cytolemma, show that the anti-CD20 bispecific single-chain antibody of anti-CD3/ has specific combination CD3 or the antigenic ability of CD20.
With certain density antibody purification and 1 * 10
6The Ramous cytomixis, 4 ℃ hatch 4h after, the centrifugal 3min of 500r/min abandons supernatant, PBS washes cell 3 times.Add 1 * 10
7The Jurkat cell, mixing, 4 ℃ hatch 4h after, can under opticmicroscope, observe the formation (Fig. 3) of rosettes.Show that the anti-CD20 single chain bispecific antibody of anti-CD3/ has the while in conjunction with CD3 and the antigenic dual specific of CD20, can effectively mediate the combination of Jurkat cell and Ramous cell.
Two. the anti-CD20 single chain bispecific antibody of anti-CD3/ is to the killing activity of human lymphocyte oncocyte
Human B cell lymphoma Ramous cell (target cell) is inoculated in 96 porocyte culture plates, every hole 100 μ l (about 1 * 10
4Individual cell), adds the anti-CD20 single chain bispecific antibody of anti-CD3/ of purifying earlier by the activity of every hole 1 μ g/ml, add the human peripheral lymphocyte (effector cell) of fresh separated again by different effect target ratios, 37 ℃, 5%CO
2Cultivate 24h, every hole adds 25 μ l MTT, 37 ℃, 5%CO
2Outwell liquid in the hole after cultivating 4h, every hole adds the acid SDS (0.1N HCl, 1%SDS) of 100 μ l, 37 ℃, 5%CO
2Hatch 4h and survey OD with microplate reader
570Value.Be calculated as follows tumor cell destruction:
Tumor cell destruction (%)=(OD
E+T-OD
E)/(1-OD
T) * 100%
In antibody concentration is under the action condition of 1 μ g/ml, and the anti-CD20 single chain bispecific antibody of anti-CD3/ increases with the rising of imitating the target ratio the killing activity of human B cell lymphoma Ramous cell.When imitating the target ratio when being 10: 1, the anti-CD20 single chain bispecific antibody of anti-CD3/ to the kill rate of human B cell lymphoma Ramous cell up to 87.3% (Fig. 4).Experimental result show the anti-CD20 single chain bispecific antibody of anti-CD3/ efficiently the Mediated Human peripheral blood lymphocyte kill and wound the human B cell lymphoma cell.
Specific embodiment of the present invention has been done detailed explanation to content of the present invention.For one of ordinary skill in the art, any conspicuous change that it is done without departing from the premise in the spirit of the present invention particularly to the replacement that is equal to of some parts, all constitutes to infringement of patent right of the present invention, with corresponding legal responsibilities.
Sequence table
＜110〉Biologic Engineering Inst., Academy of Millitary Medical Sciences of P.L.A
＜120〉the lymphadenomatous single chain bispecific antibody of a kind of anti-human B cell
<130>
<160>4
<170>PatentIn?version?3.1
<210>1
<211>121
<212>PRT
＜213〉Genus Homo, and ethnic group (Homo sapiens, human)
<400>1
Gln?Val?Gln?Leu?Gln?Gln?Pro?Gly?Ala?Glu?Leu?Val?Lys?Pro?Gly?Ala
5 10 15
Ser?Val?Lys?Met?Ser?Cys?Lys?Ala?Ser?Gly?Tyr?Thr?Phe?Thr?Ser?Tyr
20 25 30
Asn?Met?His?Trp?Val?Lys?Gln?Thr?Pro?Gly?Arg?Gly?Leu?Glu?Trp?Ile
35 40 45
Gly?Ala?Ile?Tyr?Pro?Gly?Asn?Gly?Asp?Thr?Ser?Tyr?Asn?Gln?Lys?Phe
50 55 60
Lys?Gly?Lys?Ala?Thr?Leu?Thr?Ala?Asp?Lys?Ser?Ser?Ser?Thr?Ala?Tyr
65 70 75 80
Met?Gln?Leu?Ser?Ser?Leu?Thr?Ser?Glu?Asp?Ser?Ala?Val?Tyr?Tyr?Cys
85 90 95
Ala?Arg?Ser?Thr?Tyr?Tyr?Gly?Gly?Asp?Trp?Tyr?Phe?Asn?Val?Trp?Gly
100 105 110
Ala?Gly?Thr?Thr?Val?Thr?Val?Ser?Ala
115 120
<210>2
<211>121
<212>PRT
＜213〉Genus Homo, and ethnic group (Homo sapiens, human)
<400>2
Gln?Ile?Val?Leu?Ser?Gln?Ser?Pro?Ala?Ile?Leu?Ser?Ala?Ser?Pro?Gly
5 10 15
Glu?Lys?Val?Thr?Met?Thr?Cys?Arg?Ala?Ser?Ser?Ser?Val?Ser?Tyr?Ile
20 25 30
His?Trp?Phe?Gln?Gln?Lys?Pro?Gly?Ser?Ser?Pro?Lys?Pro?Trp?Ile?Tyr
35 40 45
Ala?Thr?Ser?Asn?Leu?Ala?Ser?Gly?Val?Pro?Val?Arg?Phe?Ser?Gly?Ser
50 55 60
Gly?Ser?Gly?Thr?Ser?Tyr?Ser?Leu?Thr?Ile?Ser?Arg?Val?Glu?Ala?Glu
65 70 75 80
Asp?Ala?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Trp?Thr?Ser?Asn?Pro?Pro?Thr
85 90 95
Phe?Gly?Gly?Gly?Tyr?Lys?Leu?Glu?Ile?Lys
100 105
<210>3
<211>120
<212>PRT
＜213〉Genus Homo, and ethnic group (Homo sapiens, human)
<400>3
Gln?Val?Gln?Leu?Val?Gln?Ser?Gly?Ala?Glu?Val?Lys?Lys?Pro?Gly?Ala
5 10 15
Ser?Val?Lys?Val?Ser?Cys?Lys?Ala?Ser?Gly?Tyr?Thr?Phe?Ile?Ser?Tyr
20 25 30
Thr?Met?His?Trp?Val?Arg?Gln?Ala?Pro?Gly?Gln?Gly?Leu?Glu?Trp?Met
35 40 45
Gly?Tyr?Ile?Asn?Pro?Arg?Ser?Gly?Tyr?Thr?His?Tyr?Asn?Gln?Lys?Leu
50 55 60
Lys?Asp?Lys?Ala?Thr?Leu?Thr?Ala?Asp?Lys?Ser?Ala?Ser?Thr?Ala?Tyr
65 70 75 80
Met?Glu?Leu?Ser?Ser?Leu?Arg?Ser?Glu?Asp?Thr?Ala?Val?Tyr?Tyr?Cys
85 90 95
Ala?Arg?Ser?Ala?Tyr?Tyr?Asp?Tyr?Asp?Gly?Phe?Ala?Tyr?Trp?Gly?Gln
100 105 110
Gly?Thr?Leu?Val?Thr?Val?Ser?Ser
115 120
<210>4
<211>106
<212>PRT
＜213〉Genus Homo, and ethnic group (Homo sapiens, human)
<400>4
Asp?Ile?Gln?Met?Thr?Gln?Ser?Pro?Ser?Ser?Leu?Ser?Ala?Ser?Val?Gly
5 10 15
Asp?Arg?Val?Thr?Ile?Thr?Cys?Ser?Ala?Ser?Ser?Ser?Val?Ser?Tyr?Met
20 25 30
Asn?Trp?Tyr?Gln?Gln?Lys?Pro?Gly?Lys?Ala?Pro?Lys?Arg?Leu?Ile?Tyr
35 40 45
Asp?Thr?Ser?Lys?Leu?Ala?Ser?Gly?Val?Pro?Ser?Arg?Phe?Ser?Gly?Ser
50 55 60
Gly?Ser?Gly?Thr?Asp?Phe?Thr?Leu?Thr?Ile?Ser?Ser?Leu?Gln?Pro?Glu
65 70 75 80
Asp?Phe?Ala?Thr?Tyr?Tyr?Cys?Gln?Gln?Trp?Ser?Ser?Asn?Pro?Pro?Thr
85 90 95
Phe?Gly?Gly?Gly?Thr?Lys?Val?Glu?Ile?Lys
100 105
Claims (7)
1. lymphadenomatous single chain bispecific antibody of anti-human B cell, it is characterized in that: it is formed by connecting by anti-human B cell lymphoma CD20 single-chain antibody and anti-people CD3 single-chain antibody.
2. single chain bispecific antibody according to claim 1 is characterized in that: the connection peptides of single-chain antibody variable region is (Gly
4Ser)
3
3. single chain bispecific antibody according to claim 1 is characterized in that: the type of attachment of single-chain antibody variable region is VL
CD3-(Gly
4Ser)
3-VH
CD3-(Gly
4Ser)
3-VH
CD20-(Gly
4Ser)
3-VL
CD20
4. according to any one described single chain bispecific antibody in the claim 1 to 3, it is characterized in that: anti-human B cell lymphoma CD20 single-chain antibody has the weight chain variable region amino acid sequence shown in the sequence table SEQ ID No.1.
5. according to any one described single chain bispecific antibody in the claim 1 to 3, it is characterized in that: anti-human B cell lymphoma CD20 single-chain antibody has the light chain variable region amino acid sequence shown in the sequence table SEQ ID No.2.
6. according to any one described single chain bispecific antibody in the claim 1 to 3, it is characterized in that: anti-people CD3 single-chain antibody has the weight chain variable region amino acid sequence shown in the sequence table SEQ ID No.3.
7. according to any one described single chain bispecific antibody in the claim 1 to 3, anti-people CD3 single-chain antibody has the light chain variable region amino acid sequence shown in the sequence table SEQ ID No.4.
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|---|---|---|---|
| CNB2005100512335A CN1331888C (en) | 2005-03-02 | 2005-03-02 | Single strand double specificity anti-body resisting human B cell leucoma |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100512335A CN1331888C (en) | 2005-03-02 | 2005-03-02 | Single strand double specificity anti-body resisting human B cell leucoma |
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| Publication Number | Publication Date |
|---|---|
| CN1683409A true CN1683409A (en) | 2005-10-19 |
| CN1331888C CN1331888C (en) | 2007-08-15 |
Family
ID=35262931
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103275225A (en) * | 2013-05-10 | 2013-09-04 | 洪建� | Anti-CD31/CD146 double-specific monoclonal antibody with high affinity and application thereof |
| CN101809037B (en) * | 2007-07-31 | 2014-01-15 | 瑞泽恩制药公司 | Human antibodies to human cd20 and method of using thereof |
| CN107759694A (en) * | 2016-08-19 | 2018-03-06 | 安源医药科技（上海）有限公司 | Bispecific antibody and preparation method thereof and purposes |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113861296A (en) * | 2020-06-30 | 2021-12-31 | 广州凌腾生物医药有限公司 | Method for preparing bispecific antibody by using circular rail shaking type bioreactor |
-
2005
- 2005-03-02 CN CNB2005100512335A patent/CN1331888C/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101809037B (en) * | 2007-07-31 | 2014-01-15 | 瑞泽恩制药公司 | Human antibodies to human cd20 and method of using thereof |
| CN103275225A (en) * | 2013-05-10 | 2013-09-04 | 洪建� | Anti-CD31/CD146 double-specific monoclonal antibody with high affinity and application thereof |
| CN107759694A (en) * | 2016-08-19 | 2018-03-06 | 安源医药科技（上海）有限公司 | Bispecific antibody and preparation method thereof and purposes |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1331888C (en) | 2007-08-15 |
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