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Preparation method for monoclonal antibodies with functions of anti-tumor angiogenesis

A monoclonal antibody, anti-angiogenesis technology, applied in the field of tumor treatment, can solve the problems of low success rate, dependence and high cost

Inactive Publication Date: 2013-11-20
NANJING NORMAL UNIVERSITY
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Problems solved by technology

[0021] The purpose of the embodiment of the present invention is to provide a preparation method of anti-tumor angiogenesis monoclonal antibody, which aims to solve the existing humanization method that relies on the understanding of antibody structure informatics, which is costly, time-consuming and labor-intensive. and low success rate

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  • Preparation method for monoclonal antibodies with functions of anti-tumor angiogenesis
  • Preparation method for monoclonal antibodies with functions of anti-tumor angiogenesis
  • Preparation method for monoclonal antibodies with functions of anti-tumor angiogenesis

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[0121] The embodiment of the present invention provides a preparation method of anti-tumor angiogenesis monoclonal antibody, the preparation method of anti-tumor angiogenesis monoclonal antibody comprises the following steps:

[0122] Expression and purification of cell receptor protein VEGFR2, preparation of fully humanized phage Fab antibody library, screening of high-affinity antibodies from the phage antibody library, immobilization of purified VEGFR2 on Biacore's CM5 chip, and testing of different concentrations of AVR2 antibody and VEGFR2 the speed of association and dissociation;

[0123] Antibody library screening based on high affinity and specificity of antibodies and antigens;

[0124] Cloning of antibody genes into expressing cells, construction of engineered cell lines with high expression levels, detection and evaluation of anti-angiogenesis.

[0125] As an optimization scheme of the embodiment of the present invention, the specific method of gene cloning of the...

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Abstract

The invention discloses a preparation method for monoclonal antibodies with functions of anti-tumor angiogenesis. The preparation method comprises the following steps: expression and purification of cell receptor protein VEGFR2, preparation of completely humanized phage Fab antibody libraries, selection of antibodies with high affinity from the phage antibody libraries, immobilization of purified VEGFR2 on CM5 chips of Biacore, testing of association rates and dissociation rates of AVR2 antibodies with different concentrations and VEGFR2 respectively, selection of antibody libraries based on high affinity and high specificity of antibodies and antigens, cloning of antibody genes to expressing cells, construction of engineered cell lines with high expression level, and detection and evaluation of anti-angiogenesis. By utilizing the phage antibody library technology, the preparation method can produce humanized antibodies directly. In addition, the monoclonal antibodies are advantaged by strong drug specificity, minimal side effects and remarkable effects.

Description

technical field [0001] The invention belongs to the field of tumor treatment, in particular to a preparation method of an anti-tumor angiogenesis monoclonal antibody. Background technique [0002] Tumor growth is dependent on angiogenesis. In 1971, Dr. Folkman of Harvard Medical School found that when a tumor grows to a certain extent, it needs to establish its own blood vessel system to supply the oxygen and nutrients needed for tumor cell growth. [0003] less than 3mm 3 Tumors do not need blood vessels, and the supply of nutrients is obtained through the penetration of body fluids; when the tumor grows to a certain extent, tumor cells release some pro-angiogenic substances to induce the formation of tumor blood vessels; tumor cells obtain nutrition supply through blood vessels, and Spread of tumor cells through blood vessels. Blocking tumor blood vessel growth deprives tumor cells of oxygen and nutrients and shrinks. This is the theoretical basis for anti-angiogenic t...

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Application Information

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IPC IPC(8): C07K16/30C40B30/04
Inventor 郭志刚潘林莉高婷婷曹艳周婷
Owner NANJING NORMAL UNIVERSITY
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