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HAb18GC2 monoclonal antibody and its light and heavy chain variable area genes, coding polypeptide and use

A variable region and gene technology, applied in the fields of biotechnology and cell engineering, can solve problems such as unclear pharmacodynamic results

Active Publication Date: 2007-10-17
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

Others, such as endothelin receptor antagonists, prolyl hydroxylase inhibitors, etc. are under research, and there is no clear pharmacodynamic result yet

Method used

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  • HAb18GC2 monoclonal antibody and its light and heavy chain variable area genes, coding polypeptide and use
  • HAb18GC2 monoclonal antibody and its light and heavy chain variable area genes, coding polypeptide and use
  • HAb18GC2 monoclonal antibody and its light and heavy chain variable area genes, coding polypeptide and use

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Experimental program
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Effect test

Embodiment Construction

[0020] 1. Preparation and identification of monoclonal antibody HAb18GC2.

[0021] Immunization method: 8-10 weeks old female BALB / C mice weighing about 18 g (provided by the animal center of our school) were routinely immunized with HAb18G / CD147 extracellular domain protein, and subcutaneously injected at multiple points, 100 μg / mouse , every 2 weeks with the same dose of antigen plus incomplete Freund's adjuvant to boost immunization, intraperitoneal injection, test the titer of polyclonal antibody in mouse serum before cell fusion, and the tail vein of the higher titer will be given a booster immunization once again, the dose is the same as above. After 3 days, the splenocytes of mice immunized with HAb18G / CD147 extracellular domain protein were fused with the myeloma cell line SP2 / 0 obtained by passage in vivo at a ratio of 5:1 under the action of PEG-1500, and the fused cells were incubated in HAT selective medium cultivated in.

[0022] Establishment of hybridoma cell l...

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Abstract

The present invention relates to monoclonal antibody HAb18GC2 and its variable area gene of its heavy chain and light chain and polypeptide, and their uses in preparing therapeutic agent for reversing hepatic fibrosis etc. matrix deposition disease. The present monoclonal antibody HAb18GC2 can bind specificly with antibody HAb18G / CD147 highly expressed in human liver fibrosis tissue and with liver astrocyte and possesses function of facilitating MMPs secretion and reducing the extracellular matrix. The antibody light, heavy chain variable area genes are successfully cloned in the present invention and small molecule gene engineered antibodies of all kinds are constructed and expressed based on the gene for preparing drug for reversing liver fibrosis matrix deposition-related disease. The polypeptide coded by the present gene can crosslink with multiple effector molecule and prepare drug for reversing liver fibrosis matrix deposition-related disease.

Description

technical field [0001] The present invention relates to the field of biotechnology and cell engineering, especially monoclonal antibody HAb18GC2 and its heavy chain, light chain variable region genes, polypeptides encoded by the genes, and the above-mentioned monoclonal antibodies, light and heavy chain variable region genes, and polypeptides application. Background technique [0002] Hepatic fibrosis is a common pathological change and a necessary path for the development of various chronic liver diseases to cirrhosis. Chronic liver disease mainly includes various types of viral hepatitis, especially hepatitis C and hepatitis B. There are more than 280 million HbsAg carriers worldwide. my country is a high prevalence area of ​​hepatitis B, with 130 million HbsAg carriers and more than 30 million hepatitis B patients. 25-40% of hepatitis B patients lead to liver cirrhosis and liver cancer, and the death toll is 400,000 per year. It has become a serious disease in my countr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C12N5/18C12N15/13A61K39/395A61P1/16
Inventor 陈志南张思河杨向民汪莉李郁张征
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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