Method for producing anti-osteopontic antibody medicine and use

A technology for osteopontin and production methods, which can be used in the fields of antibodies, drug combinations, cardiovascular system diseases, etc., can solve the problems of laboratory research and drug development that have not been publicly reported or retrieved, and can inhibit cell adhesion and prevent Adhesion and inhibition of calcium salt deposition

Inactive Publication Date: 2002-06-05
HEBEI MEDICAL UNIVERSITY
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  • Abstract
  • Description
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Problems solved by technology

Using soluble adhesion molecules or anti-adhesion molecule antibodies to block the interaction between vascular smooth muscle cells and extracellular matrix can inhibit the migration of vascular smooth muscle cells. However, at present, the research and development of soluble adhesion molecules and their antibodies in my country It is still in a backward state, and there are no public reports on laboratory research and drug development
Novel searches at home and abroad have found any research reports on using OPN as a tissue growth promoter and using anti-OPN single-chain antibodies or small molecule peptide antagonists to block the migration and proliferation of vascular smooth muscle cells, and no one has screened from the phage antibody library. Humanized antibody OPN single chain antibody and screening of OPN antagonists from phage random short peptide library for patent protection

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0013] Example 1: Production of human anti-OPN single chain antibody.

[0014] 1. Construction of phage human antibody library

[0015] Peripheral blood mononuclear cells from patients after PTCA were isolated, sensitized in vitro with rat OPN, transformed with EBV, enriched B cells secreting specific antibodies, and extracted total RNP. The heavy chain and light chain variable region gene fragments of the antibody and the DNA linker encoding the pentapeptide (Gly4Ser) 3 were assembled together by recombinant PCR to construct a phage antibody library.

[0016] 2. Use human OPN to screen anti-OPN single-chain antibodies from the phage antibody library, soak the nitrocellulose membrane in (70ug / ml) human OPN solution, place it at room temperature and dry it, put it in a polypropylene plastic plate, and take 2ml of phage suspension The solution was added to the plate, shaken at room temperature for 1 to 2 hours, washed with PBS to remove unbound phages, replaced with 2ml elution...

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Abstract

The present invention relates to the field of biological preparation technology. By means of bioengineering process, the humanized anti-osteopontin single chain antibody, anti-osteopontin monoclonal antibody and anti-osteopontin polyclonal antibody are produced and made into various medicament forms and are used in preventing and treating vascular re-stenosis, atheoscleorsis and osteoporosis. The present invention has the features of novel materials, unique and reasonable production process, high curative effect and no toxic side-effect, and wide application prospect.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals and medicines for preventing and treating blood vessel restenosis, atherosclerosis and osteoporosis. Background technique [0002] Cardiovascular disease is one of the most serious diseases that endanger human life and health, and its incidence rate accounts for the first of all kinds of diseases. Coronary heart disease is the main cause of death of such patients. Percutaneous transluminal coronary angioplasty (PTCA) is one of the important methods for the treatment of coronary heart disease, and millions of people around the world receive this treatment every year. However, since 30%-50% of patients have restenosis within 6 months after operation, there is no drug with definite curative effect yet. Placement of intravascular stents is the main measure to prevent restenosis. Stent implantation can prevent arterial elastic recoil and prevent vessel wall remodeling, thereby reducing the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P9/10C12P21/00
Inventor 韩梅温进坤
Owner HEBEI MEDICAL UNIVERSITY
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