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HAb18Gedomab 1 monoclonal antibody and light and heavy chain variable area genes, coding polypeptide and use

A monoclonal antibody, variable region technology, applied in the field of biotechnology and cell engineering, can solve the problem of no clear pharmacodynamic results and so on

Active Publication Date: 2009-04-22
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Claims
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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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  • HAb18Gedomab 1 monoclonal antibody and light and heavy chain variable area genes, coding polypeptide and use
  • HAb18Gedomab 1 monoclonal antibody and light and heavy chain variable area genes, coding polypeptide and use
  • HAb18Gedomab 1 monoclonal antibody and light and heavy chain variable area genes, coding polypeptide and use

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Experimental program
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Embodiment Construction

[0018] 1. Preparation and identification of monoclonal antibody secreted by HAb18Gedomab1.

[0019] 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-1000, and the fused cells were incubated in HAT selective medium cultivated in.

[0020] Establishment of...

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Abstract

The present invention immunizes mouse with HAb18G / CD147 extra-cellular region protein to prepare specific hybrid tumor cell strain HAb18Gedomab1, obtain monoclonal antibody HAb18Gedomab1, which can combine specifically to HAb18G / CD147 expressed effectively in human liver fibrosis tissue and liver stellate cell and possesses functions of promoting MMPs secretion and degrades extra-cellular matrix. The antibody may be used in preparing medicine for reversing liver fibrosis and other matrix deposition related diseases. The present invention clones the light and heavy chain variable area genes successively. Several forms of small molecular gene engineering antibody are constituted and expressed based on the genes and gene engineering method, and the polypeptide coded based on the genes is cross-linked with several effect factors, so as to prepare medicine for reversing liver fibrosis and other matrix deposition related diseases.

Description

technical field [0001] The present invention relates to the fields of biotechnology and cell engineering, especially the monoclonal antibody secreted by HAb18Gedomab1 and its heavy chain, light chain variable region genes, polypeptides encoded by the genes, and the above-mentioned monoclonal antibodies and light and heavy chain variable regions Application of genes and peptides. 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 seri...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/13C12N5/12C07K16/18A61K39/00A61P1/16
Inventor 陈志南邢金良张思河杨向民汪莉叶卉李别虎
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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