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
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[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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