CN107748258A - A kind of ELISA detection method of Fc γ RII acceptors - Google Patents
A kind of ELISA detection method of Fc γ RII acceptors Download PDFInfo
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- CN107748258A CN107748258A CN201710616081.1A CN201710616081A CN107748258A CN 107748258 A CN107748258 A CN 107748258A CN 201710616081 A CN201710616081 A CN 201710616081A CN 107748258 A CN107748258 A CN 107748258A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Abstract
A kind of ELISA detection method of claimed Fc γ RII acceptors, the step of it is included in the coating step that Fc γ RII CD32 are coated with ELISA Plate, closing, sample preparation, secondary antibody are incubated, substrate colour developing, wherein, the sample preparation step is：After anti-VEGF mAb mixes with antigen, a hour formation immune complex is stood；The mol ratio of anti-VEGF mAb and antigen is 0.1~10:Between 1；Immune complex progress gradient dilution is added to be coated with Fc γ RII CD32 ELISA Plate and is incubated.This method is initially formed the situation that immune complex is combined with acceptor again using the antibody in ELISA method analogue body, the form of receptor antibody secondary antibody is developed into the form of receptor immunocomplexes secondary antibody, improve anti-VEGF mAb and Fc γ RIIA (H131), Fc γ RIIA (R131), Fc γ RIIB binding activity, sensitivity, the stability of detection method are improved, complete four parameter curve can be obtained.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of ELISA detection method of Fc γ RII (CD32) acceptor.
Background technology
VEGF (vascular endothelial growth factor, VEGF), is blood vessel endothelium
The HBGF of cell-specific, can in vivo induction of vascular it is newborn.VEGF also may be used by different tumor cell secretions
Developmental normal cell and tissue in express, in vivo adjust blood vessel permeability, be vascularization during it is a variety of
Cell provides a network of fibers；The degraded of matrix and propagation, migration, motion and the blood vessel of endothelial cell can be promoted in vitro
The formation of chamber spline structure, and the endothelial progenitor cells of derived from bone marrow can be mobilized.VEGF be the effect of induced tumor vascularization it is most strong,
Specific highest angiogenesis factor.So in theory, employ different approach to intervene or suppress vascular endothelial growth
The factor can suppress the growth of tumour.
Therapeutic monoclonal antibodies have turned into a kind of and have tackled cancer and the critical treatment means of autoimmune disease.Monoclonal antibody
Effect depends not only upon its antibody binding domain (Fab), while its crystallizable region (Fc) also plays an important role.Fab energy
It is enough specifically to identify tumor associated antigen (TAA), so as to regulate and control the related downstream signaling pathways of TAA.The Fc regions energy of IgG molecules
Play a series of biological functions and further lift monoclonal antibody effect.Fc can be mainly expressed all kinds of with reference to Fc γ Rs, Fc γ Rs
On immune leucocyte, when this kind of cell is recruited and is activated, Antibody -dependent cell cytotoxicity can be triggered to act on (ADCC)
And the phagocytosis (ADCP) of antibody-dependant cell mediation, so as to remove target tumor.Fc can also combine serum and mend
Body molecule (C1q), then form membrane attack complex (MAC) and remove aim cell, this process is referred to as the thin of Complement Dependent
Cytotoxicity (CDC).Fc regions can also combine newborn Fc acceptors (FcRn) in a manner of PH is relied on, and avoid antibody by cell
Endosome is degraded, so as to extend the half-life period of monoclonal antibody.
There are the Fc acceptors of four class IgG molecules in human body：FcγRI(CD64),FcγRII(CD32),FcγRIII
And FcRn (CD16).Wherein Fc γ RI, Fc γ RIIa, Fc γ RIIc and Fc γ RIIIa can pass through its IMAM activation sequence
Immune effector cell is stimulated to activate, so as to trigger ADCC and ADCP.Corresponding, Fc γ RIIb include an ITIM and suppress sequence,
So as to suppress inflammatory reaction.Fc γ RIIb are that only one plays the Fc acceptors for suppressing function.It generally with other activating Fcs
γ Rs are expressed on identical cell, and negative-feedback regu- lation effect, therefore Fc are played for Fc γ RIIa and Fc γ RIIIa signal
γ RIIb can be applied to the treatment of autoimmune disease and anaphylactic disease.Expressed in addition, Fc γ RIIb are only ones in B cell
On Fc acceptors, therefore the B cell of antigen-specific can identify the compound of autoantigen and treatment antibody by Fc γ RIIb,
So as to play immunosuppressive action.So Fc γ RIIb can be as the critical checkpoints of humoral immune reaction, in lupus erythematosus
Etc. (SLE) played a role in disease.
Fc γ RII (CD32) basic structure：40kDa wears membrane glycoprotein, belongs to Ig superfamily members, and after birth outskirt has 2
C2 structures, Genetic food is in 1q23~24.Fc γ RII (CD32) II and monomer human IgG1, IgG3, IgG4 are combined into low
Affinity, Kd5 × 10-7M.Fc γ RII (CD32) are expressed in except exo-erythrocytic other haemocytes, molecule amount：20000~
40000/ each cell.Different with function according to DNA sequence dna, Fc γ RII (CD32) can be divided into three kinds of form D c γ RIIA, Fc γ
RIIB, Fc γ RIIC, multi-form Fc γ RII difference essentially consist in the structure difference of cytoplasmic domain.GM-CSF is to acidophil granules
The activation of cell is mainly by Fc γ RII mediations.Research Fc γ Rs and IgG combination for understand immunity of organism and
Antibody drug research and development have great importance.
The content of the invention
The technical problems to be solved by the invention are for anti-VEGF mAb and Fc γ RIIA (H131), Fc γ RIIA
(R131), the binding ability of Fc γ RIIB acceptors is very weak, and it is bent to form complete four parameter with common ELISA method
Line.Sandwich method ELISA is extended by the invention, and the form of receptor-antibody-secondary antibody is developed into acceptor-immune
The form of compound-secondary antibody, it is intended to improve antibody and Fc γ RIIA (H131), Fc γ RIIA (R131), Fc γ RIIB combination
Activity, sensitivity, the stability of ELISA detection method are improved, complete four parameter curve can be obtained.
In order to solve the above-mentioned technical problem, technical scheme provided by the invention is that one kind ELISA method detects anti-vegf
Monoclonal antibody medicine and the method for Fc γ RII (CD32) binding activity, it is included in the coating that Fc γ RII-CD32 are coated with ELISA Plate
The step of step, closing, sample preparation, secondary antibody incubation, substrate colour developing, and the sample preparation step is：Anti-VEGF mAb with
After antigen mixes, a hour formation immune complex is stood.Compared with the ELISA method of routine, this method can obtain completely
Four parameter curves (present completely upper lower platform), and there is good repeatability.
In currently preferred technical scheme, the Fc γ RII-CD32 are selected from Fc γ RIIA (H131), Fc γ RIIA
(R131), one kind in Fc γ RIIB.
In currently preferred technical scheme, the mol ratio of anti-VEGF mAb and antigen is 0.1~10:Between 1；It is preferred that
Ground, the mol ratio of the anti-VEGF mAb and antigen is 0.5~5:Between 1, it is highly preferred that anti-VEGF mAb medicine and antigen
Mol ratio is 1~3:Between 1.
In currently preferred technical scheme, gradient dilution is carried out to immune complex, addition has been coated with Fc γ RII-
It is incubated on CD32 ELISA Plate
Fc γ RIIA (H131), Fc γ RIIA (R131), Fc are coated with currently preferred technical scheme, on ELISA Plate
More than one or both of γ RIIB mixture.
The anti-VEGF mAb includes but is not limited to Avastin BEVACIZUMAB (Avastin).
In currently preferred technical scheme, the molecular formula and molecular weight of the anti-VEGF mAb are：
C6638H10160N1720O2108S44149kDa, its light-chain amino acid sequence are as follows：
DIQMTQSPSS LSASVGDRVT ITCSASQDIS NYLNWYQQKP GKAPKVLIYF TSSLHSGVPS
RFSGSGSGTD FTLTISSLQP EDFATYYCQQ YSTVPWTFGQGTKVEIKRTV AAPSVFIFPP SDEQLKSGTA
SVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQG
LSSPVTKSFN RGEC；
The amino acid sequence of heavy chain is：
EVQLVESGGG LVQPGGSLRL SCAASGYTFT NYGMNWVRQA PGKGLEWVGW INTYTGEPTY
AADFKRRFTF SLDTSKSTAY LQMNSLRAEDTAVYYCAKYP HYYGSSHWYF DVWGQGTLVT VSSASTKGPS
VFPLAPSSKS TSGGTAALGC LVKDYFPEPV TVSWNSGALT SGVHTFPAVLQSSGLYSLSS VVTVPSSSLG
TQTYICNVNH KPSNTKVDKK VEPKSCDKTH TCPPCPAPEL LGGPSVFLFP PKPKDTLMIS RTPEVTCVVV
DVSHEDPEVKFNWYVDGVEV HNAKTKPREE QYNSTYRVVS VLTVLHQDWL NGKEYKCKVS NKALPAPIEK
TISKAKGQPR EPQVYTLPPS REEMTKNQVSLTCLVKGFYP SDIAVEWESN GQPENNYKTT PPVLDSDGSF
FLYSKLTVDK SRWQQGNVFS CSVMHEALHN HYTQKSLSLS PGK。
A kind of recombinant humanized Anti-X activity of the present invention is a kind of therapeutic monoclonal antibodies product, and it can
VEGF (vascular endothelial growth factor, VEGF) is specifically bound, and is blocked
VEGF biological activity, to suppress angiogenesis, so as to reach antitumor action.Chinese storehouse is passed through using recombinant DNA technology
The recombinant humanized monoclonal IgG1 antibody of mouse ovary (Chinese hamster ovary, CHO) cell expression system production,
The mouse source monoclonal antibody complementary determining region sequence with reference to people VEGF comprising 93% Human monoclonal antibody sequence and 7%, heavy chain is by 453
Amino acid is formed, and light chain is made up of 214 amino acid, and molecular weight is about 149kDa.
The present invention have studied the combination of Fc γ RIIA (H131), Fc γ RIIA (R131), Fc γ RIIB and anti-VEGF mAb
Method.In recent years, biomembrane interference technique (biolayer interferomeory, BLI), biacore technologies etc. are used for Fc
The measure of γ Rs binding activity, although simple and efficient, expensive large scale equipment is needed, be unfavorable for common lab, small
The technical research of type biotech firm and the tracking and monitoring of product quality, and simply detection monomer IgG molecules and Fc γ Rs, do not have
The situations of analog antibody IgG in vivo.And enzyme-linked immunosorbent assay (ELISA) method, because its is simple to operate, quick, quick
Perceptual height, high specificity, equipment requirement are simple, therefore in laboratory extensive use.Anti-VEGF mAb and Fc γ in the present invention
RIIA (H131), Fc γ RIIA (R131), Fc γ RIIB receptor-binding activity detection methods are to be based on ELISA method, analogue body
Interior antibody is initially formed the situation that immune complex is combined with acceptor again, utilizes parallel curves (four parameter curves) model determination phase
To activity.During method is developed, it is found that the binding ability of monomer IgG monoclonal antibodies and Fc γ RIIA, Fc γ RIIB is very weak, no
Complete four parameter curve can be made., could well and Fc after monomer IgG and VEGF are incubated to form immune complex altogether
γ RIIA, Fc γ RIIB are combined, and form complete four parameter curve.This with internal situation be it is consistent, when antibody not with
When immunogene combines, internal immune response can not be caused.The immune anti-of body can just be caused after only being combined with immunogene
Should.Therefore this method simulates internal situation well.
At present, domestic detection anti-VEGF mAb and Fc γ RIIA (H131), Fc γ RIIA (R131), Fc γ RIIB acceptor knots
Close mainly using biomembrane interference technique (biolayer interferomeory, BLI), biacore technologies, it is impossible to well
Situation in analogue body.The domestic document report for not detecting antibody and Fc γ RII with ELISA method also at present.This method is adopted
The situation that immune complex combined with acceptor again is initially formed with the antibody in ELISA method analogue body, by receptor-antibody-secondary antibody
Form develop into the form of acceptor-immune complex-secondary antibody, improve anti-VEGF mAb and Fc γ RIIA (H131), Fc γ
RIIA (R131), Fc γ RIIB binding activity, sensitivity, the stability of detection method are improved, complete four ginseng can be obtained
Number curve.This method can evaluate the uniformity of different batches anti-VEGF mAb and the anti-VEGF mAb of different manufacturers production,
Can have extensively in the tracking and monitoring of antibody drug research and development and product quality as the indirect indexes of evaluation monoclonal antibody pharmacokinetics
Application.
Brief description of the drawings
Fig. 1 is Fc γ RII and anti-VEGF mAb combination detection method schematic diagram.
Fig. 2 is Fc γ RIIA (H131) and anti-VEGF mAb binding curve figure.
Fig. 3 is Fc γ RIIA (R131), Fc γ RIIB and anti-VEGF mAb binding curve figure.
Fig. 4 A are the light-chain amino acid sequence figure of anti-VEGF mAb；Fig. 4 B are the heavy chain amino acid sequence of anti-VEGF mAb
Figure.
Embodiment
Such scheme is described further below in conjunction with specific embodiment.It should be understood that these embodiments are to be used to illustrate
The present invention and be not limited to limit the scope of the present invention.The implementation condition used in embodiment can be done according to the condition of specific producer
Further adjustment, unreceipted implementation condition is usually the condition in normal experiment.
Introduce and summarize
The present invention by way of example rather than provides the mode of limitation to illustrate.It should be noted that in present disclosure
Described " one " or " one kind " embodiment is not necessarily referring to same embodiment, and refers at least a kind of.
Various aspects of the invention are described below.However, as will be readily apparent to one of skill in the art, can
Implement the present invention according to the only some or all of aspects of the present invention.For purposes of illustration, provide herein specific numbering, material and
Configuration, enables one to thoroughly understand the present invention.However, be evident that for those of skill in the art,
The present invention can be implemented without concrete details.In other examples, not make the present invention is obscure many institutes have been omitted or simplified
Known feature.
Various operations are described successively as multiple discrete steps, and with most helpful in the side for understanding the present invention
Formula illustrates；However, in-order description should not be construed as to imply that these operations are necessarily dependent on order.
Reactant according to type species is illustrated to various embodiments.To show for those of skill in the art and
It is clear to, any number of different types of reactant can be used to implement for the present invention, and be more than those for the purpose of illustration
And the reactant provided herein.In addition, also it is evident that, the invention is not limited in any specific mixing is shown
Example.
Experiment material and equipment
Basic model eddy mixer, ELIASA (producer：Molecular Devices, model：Spectra Max M2e)、
ELISA Plate (is purchased from：Corning), microwell plate constant temperature oscillator, board-washing machine (are purchased from：Thermo)
Experiment reagent：
Phosphate buffer (PBS solution), sample diluting liquid/washing lotion, confining liquid, nitrite ion TMB, terminate liquid (1M sulphur
Acid), antigen rhVEGF165, Fc γ RIIA (H131), Fc γ RIIA (R131), Fc γ RIIB
Test sample：All test samples provide by TOT Biopharm Co., Ltd..
Embodiment 1：Fc γ RIIA (H131) and anti-VEGF mAb associated methods
Sample preparation：
(1) anti-VEGF mAb Avastin and reference material are diluted to 30 μ g/ml concentration respectively with sample diluting liquid.
(2) the 30 μ g/ml μ l of sample 200 are taken, add the VEGF that concentration is 0.5mg/ml thereto165, anti-vegf list is made
It is anti-：VEGF mol ratio is 1:Mixed liquor between 1, it is stored at room temperature after mixing 1~2 hour.
(3) by above-mentioned 30 μ g/ml 8~11 concentration gradients of concentration mixed liquor gradient dilution.
Experimental procedure：
(1) each hole of ELISA Plate is added with the μ g/ml of 100 μ l 2.5 Fc γ RIIA (H131) coating buffer solutions；Use sealed membrane
After sealing, 16-20 hours are incubated under the conditions of being placed in 2-8 DEG C.
(2) after taking-up ELISA Plate discards coating buffer, 300 μ l washing lotions (wash buffer) board-washings two are added per hole with board-washing machine
Time, plate is patted dry on filter paper.
(3) 100 μ l confining liquids are added in each hole, after being sealed with sealed membrane, 200rpm is in micropore at ambient temperature
Shaking is incubated 1-2h in plate oscillator.
(4) microwell plate is removed from micropore plate oscillator, topples over content, patted dry, add 300 μ l washing lotions per hole with board-washing machine
(wash buffer) is cleaned, and is patted dry, is repeated 4 times.
(5) for the microwell plate being coated with, each 100 μ l of need testing solution of gradient dilution are added in working hole respectively.
After being sealed with sealed membrane, 200rpm shakings at ambient temperature are incubated 1-2h.
(6) microwell plate is removed from micropore plate oscillator, topples over content, patted dry, add 300 μ l washing lotions per hole with board-washing machine
(wash buffer) is cleaned, and is patted dry, is repeated 6 times.
(7) the ELIAS secondary antibody dilutions of 100 μ l 5000~10000 times of dilutions are added in every hole, after being sealed with sealed membrane,
200rpm shakings at ambient temperature are incubated 1~2h.
(8) microwell plate is removed from micropore plate oscillator, topples over content, patted dry, add 300 μ l washing lotions per hole with board-washing machine
(wash buffer) is cleaned, and is patted dry, is repeated 6 times.
(9) 100 μ l TMB nitrite ions, sealing are added in each working hole respectively, room temperature lucifuge is incubated 10~20 minutes, work
Make hole and blueness occur.
(10) add 100 μ l terminate liquid respectively per hole, rap microwell plate mixing, enzyme reaction terminates, and originally shows the hole of blueness
Will yellowing.
(11) in terminating reaction half an hour, light absorption value is surveyed in 450nm wavelength, produces the dosage of standard liquid and sample solution
Effect curve.
(12) carried out curve fitting calculating using ELIASA software.
Embodiment 2：Fc γ RIIA (R131), Fc γ RIIB and anti-VEGF mAb associated methods
Sample preparation：
(1) anti-VEGF mAb Avastin and reference material are diluted to 50 μ g/ml concentration respectively with sample diluting liquid.
(2) the 50 μ g/ml μ l of sample 200 are taken, add the VEGF that concentration is 0.3mg/ml thereto165, anti-vegf list is made
It is anti-：VEGF mol ratio is 3:Mixed liquor between 1,1-2 hours are stored at room temperature after mixing.
(3) by 8~11 concentrations of above-mentioned 50 μ g/ml mixed liquors gradient dilution.
Experimental procedure：
(1) each hole of ELISA Plate is added with the μ g/ml of 100 μ l 5 Fc γ RIIA (R131) or Fc γ RIIB coating buffer solution；
After being sealed with sealed membrane, it is incubated 16 hours under the conditions of being placed in 5 DEG C.
(2) after taking-up ELISA Plate discards coating buffer, 300 μ l washing lotions (wash buffer) board-washings two are added per hole with board-washing machine
Time, plate is patted dry on filter paper.
(3) 100 μ l confining liquids are added in each hole, after being sealed with sealed membrane, 200rpm is in micropore at ambient temperature
Shaking is incubated 2h in plate oscillator.
(4) microwell plate is removed from micropore plate oscillator, topples over content, patted dry, add 300 μ l washing lotions per hole with board-washing machine
(washbuffer) clean, pat dry, be repeated 4 times.
(5) for the microwell plate being coated with, each 100 μ of need testing solution of 3 times of gradient dilutions is added in working hole respectively
L, after being sealed with sealed membrane, 200rpm shakings at ambient temperature are incubated 2h.
(6) microwell plate is removed from micropore plate oscillator, topples over content, patted dry, add 300 μ l washing lotions per hole with board-washing machine
(wash buffer) is cleaned, and is patted dry, is repeated 6 times.
(7) the ELIAS secondary antibody dilutions of 100 μ l 5000~10000 times of dilutions are added in every hole, after being sealed with sealed membrane,
200rpm shakings at ambient temperature are incubated 1h.
(8) microwell plate is removed from micropore plate oscillator, topples over content, patted dry, add 300 μ l washing lotions per hole with board-washing machine
(wash buffer) is cleaned, and is patted dry, is repeated 6 times.
(9) 100 μ l TMB nitrite ions, sealing are added in each working hole respectively, room temperature lucifuge is incubated 20~30 minutes, work
Make hole and blueness occur.
(10) add 100 μ l terminate liquid respectively per hole, rap microwell plate mixing, enzyme reaction terminates, and originally shows the hole of blueness
Will yellowing.
(11) in terminating reaction half an hour, light absorption value is surveyed in 450nm wavelength, produces the dosage of standard liquid and sample solution
Effect curve.
(12) carried out curve fitting calculating using SOFTMAX softwares.
Specific embodiment described above is only the preferred embodiment of the present invention, it is noted that for the art
For those of ordinary skill, under the premise without departing from the principles of the invention, some improvement or replacement can also be made, these improvement
Or replace and should also be as being considered as protection scope of the present invention.
Claims (7)
1. a kind of ELISA detection method of Fc γ RII acceptors, it is characterised in that it, which is included on ELISA Plate, is coated with Fc γ RII-
The step of CD32 coating step, closing, sample preparation, secondary antibody incubation, substrate develop the color, wherein, the sample preparation step is：
After anti-VEGF mAb mixes with antigen, a hour formation immune complex is stood.
2. detection method according to claim 1, it is characterised in that the mol ratio of the anti-VEGF mAb and antigen exists
0.1~10:Between 1.
3. detection method according to claim 1, it is characterised in that the Fc γ RII-CD32 are selected from Fc γ RIIA-
One kind in H131, Fc γ RIIA-R131, Fc γ RIIB.
4. detection method according to claim 1, it is characterised in that gradient dilution is carried out to the immune complex, added
Enter to be coated with and be incubated on Fc γ RII-CD32 ELISA Plate.
5. detection method according to claim 1, it is characterised in that the mol ratio of the anti-VEGF mAb and antigen exists
0.5~5:Between 1.
6. detection method according to claim 1, it is characterised in that Fc γ RIIA-H131, Fc γ are coated with ELISA Plate
Mixture more than one or both of RIIA-R131, Fc γ RIIB.
7. detection method according to claim 1, it is characterised in that the anti-VEGF mAb is Avastin.
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| CN101268372A (en) * | 2005-07-21 | 2008-09-17 | 根马布股份公司 | Potency assays for antibody drug substance binding to FC receptor |
| US20130315907A1 (en) * | 2012-05-22 | 2013-11-28 | Regeneron Pharmaceuticals, Inc. | Vegf-a121 assay |
| CN103460048A (en) * | 2011-02-22 | 2013-12-18 | 伯恩哈德－诺策热带医学研究所 | Detection of antibodies using an improved immune complex (IC) ELISA |
| WO2017048699A2 (en) * | 2015-09-14 | 2017-03-23 | Galaxy Biotech Llc | Highly potent monoclonal antibodies to angiogenic factors |
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2017
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1763097A (en) * | 1999-01-15 | 2006-04-26 | 杰南技术公司 | Polypeptide variants with altered effector function |
| CN101268372A (en) * | 2005-07-21 | 2008-09-17 | 根马布股份公司 | Potency assays for antibody drug substance binding to FC receptor |
| CN103460048A (en) * | 2011-02-22 | 2013-12-18 | 伯恩哈德－诺策热带医学研究所 | Detection of antibodies using an improved immune complex (IC) ELISA |
| US20130315907A1 (en) * | 2012-05-22 | 2013-11-28 | Regeneron Pharmaceuticals, Inc. | Vegf-a121 assay |
| WO2017048699A2 (en) * | 2015-09-14 | 2017-03-23 | Galaxy Biotech Llc | Highly potent monoclonal antibodies to angiogenic factors |
Non-Patent Citations (3)
| Title |
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| ADAM WALKER ET AL: "Novel Interaction Mechanism of a Domain Antibody-based Inhibitor of Human Vascular Endothelial Growth Factor with Greater Potency than Ranibizumab and Bevacizumab and Improved Capacity over Aflibercept", 《THE JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
| DOUGLAS A. MACDONALD ET AL: "Aflibercept exhibits VEGF binding stoichiometry distinct from bevacizumab and does not support formation of immune-like complexes", 《ANGIOGENESIS》 * |
| ROBERT L.SHIELDS, ET AL.: "High Resolution Mapping of the Binding Site on Human IgG1 for FcγRI, FcγRII, FcγRIII, and FcRn and Design of IgG1 Variants with Improved Binding to the FcγR.", 《THE JOURNAL OF BIOLOGICAL CHEMISTRY》 * |
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Application publication date: 20180302 |
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