Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Biological activity analysis method of recombinant anti-VEGFR2 monoclonal antibody and application thereof

A monoclonal antibody, biologically active technology, applied in the field of biological analysis, to achieve the effect of high accuracy, high sensitivity and wide application range

Pending Publication Date: 2020-04-17
MAB VENTURE BIOPHARM CO LTD
View PDF10 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The application value of recombinant anti-VEGFR2 monoclonal antibody is huge, and its related research must involve the evaluation of its biological activity. At present, there is no specific method for its biological activity analysis. Therefore, the completion of the recombinant anti-VEGFR2 monoclonal antibody The development of biological cell activity methods for antibodies is very meaningful

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Biological activity analysis method of recombinant anti-VEGFR2 monoclonal antibody and application thereof
  • Biological activity analysis method of recombinant anti-VEGFR2 monoclonal antibody and application thereof
  • Biological activity analysis method of recombinant anti-VEGFR2 monoclonal antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] This embodiment provides a method for analyzing the biological activity of a recombinant anti-VEGFR2 monoclonal antibody, comprising the following steps:

[0051] (1) Take HUVEC cells in the logarithmic growth phase, digest the cells according to the cell subculture process, wash the digested cells with ECM complete medium and centrifuge, then resuspend the cells with ECM complete medium to make HUVEC cells Cell suspension, mix well, take 0.3mL for cell counting, and do 2 parallel operations. The number of living cells and cell viability were determined according to the mean value of the two cell counts. Then adjust the viable cell density to 0.5×10 with ECM complete medium 5 Cells / mL, add 100 μL / well into 96-well cell culture plate. Place the 96-well cell culture plate at 37 °C, 5% CO 2 Cultured in a constant temperature incubator for 6 h. Discard the supernatant, add 0.1% FBS ECM analysis medium at 100 μL / well, seal the periphery with PBS, and store at 37°C, 5% CO...

Embodiment 2

[0056] This embodiment provides a method for analyzing the biological activity of a recombinant anti-VEGFR2 monoclonal antibody, comprising the following steps:

[0057] (1) Take HUVEC cells in the logarithmic growth phase, digest the cells according to the cell subculture process, wash the digested cells with ECM complete medium and centrifuge, then resuspend the cells with ECM complete medium to make HUVEC cells Cell suspension, mix well, take 0.3mL for cell counting, and do 2 parallel operations. The number of living cells and cell viability were determined according to the mean value of the two cell counts. Then adjust the viable cell density to 1×10 with ECM complete medium 5 Cells / mL, add 100 μL / well into 96-well cell culture plate. Place the 96-well cell culture plate at 37 °C, 5% CO 2 Cultured in a constant temperature incubator for 6 h. Discard the supernatant, add 0.1% FBS ECM analysis medium at 100 μL / well, seal the periphery with PBS, and store at 37°C, 5% CO ...

Embodiment 3

[0062] This embodiment provides a biological activity analysis method of a recombinant anti-VEGFR2 monoclonal antibody. The difference between the analysis method and Example 1 is that VEGF165 is diluted to 0.4 μg / mL with the sample diluent in step (2), and the other are all constant.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a biological activity analysis method of a recombinant anti-VEGFR2 monoclonal antibody and an application thereof. The biological activity analysis method comprises the following steps: co-culturing a recombinant anti-VEGFR2 monoclonal antibody, human umbilical vein endothelial cells and a VEGF reagent; adding a CCK8 chromogenic solution for continued culture; reading a light absorption value; carrying out 4-parameter fitting according to the dose-effect relationship between the concentration of the recombinant anti-VEGFR2 monoclonal antibody and the light absorption value, and carrying out biological activity evaluation according to an EC50 value of curve fitting. The method is a biological activity analysis method special for a recombinant anti-VEGFR2 monoclonalantibody, has the advantages of simple operation, high sensitivity, high accuracy and wide application range, can be used for detecting recombinant anti-VEGFR2 monoclonal antibodies with different titer levels, and can be used for detecting the biological activity of intermediates, stock solutions or finished products of the recombinant anti-VEGFR2 monoclonal antibodies.

Description

technical field [0001] The invention belongs to the technical field of biological analysis, and in particular relates to a biological activity analysis method of a monoclonal antibody and an application thereof, in particular to a biological activity analysis method of a recombinant anti-VEGFR2 monoclonal antibody and an application thereof. Background technique [0002] VEGFR2 is highly expressed on tumor-associated endothelial cells. When VEGF binds to VEGFR2 on tumor cell-associated endothelial cells, it initiates a signal cascade reaction, causing endothelial cell proliferation and metastasis, and increasing the permeability of blood vessel walls, which will lead to angiogenesis and provide sufficient oxygen and nutrients for tumor occurrence and development . Anti-VEGFR2 recombinant fully human monoclonal antibody can specifically block the interaction between vascular endothelial growth factor receptor 2 (VEGFR-2) and its ligand VEGF, inhibit the activation of VEGFR2 ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577C12N5/071
CPCG01N33/577C12N5/069C12N2501/165
Inventor 王少雄吕品
Owner MAB VENTURE BIOPHARM CO LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com