[原著]Estimation of rabbit fibrinopeptide B using a monoclonal antibody
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We produced a monoclonal antibody against rabbit fibrinopeptide B (FPB) and established an immunological assay system to estimate it. Rabbit FPB was purified from the supernatant of fibrin clot prepared with rabbit fibrinogen and bovine thrombin. The purified rabbit FPB was coupled to keyhole limpet hemocyanin (KLH), and BALB/c mice were immunized with the resultant FPB-KLH. The spleen cells of immunized mice were hybridized with P3-X63-Ag8-U1 myeloma cells. As a result, one hybridoma (IA12/4A2) was selected, which secreted an antibody against rabbit FPB. The monoclonal antibody obtained belonged to the IgM class and its light chain consisted of a $ \kappa $-chain. The monoclonal antibody reacted not only with rabbit FPB but also rabbit fibrinogen, however it did not react with rabbit fibnnopeptide A. Utilizing this monoclonal antibody, we attempted to develop a competitive enzyme-linked immunoassay for estimating purified FPB. The working range of FPB varied between 5 and 50 $ \mu $g/ml.
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