ANTI-HUMAN VITRONECTIN/THROMBIN/ANTITHROMBIN III COMPLEX MONOCLONAL ANTIBODY, HYBRIDOMA AND IMMUNOASSAY

摘要

PURPOSE: To obtain the subject new antibody for diagnosing blood coagulation blood coagulation sthenia, accentuation, specifically reacting with vitronectin/ thrombin/antithrombin III complex (VTAT), not reacting with each liberated material, capable of measuring VTAT in high sensitivity. ;CONSTITUTION: Vitronectin is reacted with thrombin and antithrombin III in 1M tris hydrochloric acid buffer solution (pH7.5) containing 0.9% NaCl at 37°C for 30 minutes and purified by molecular sieve chromatography to obtain VTAT. Then VTAT and an adjuvant are administered to the abdominal cavity of a mouse and the mouse is immunized. After the final immunization, splenic cells are collected, fused with mouse myeloma cells and selectively cultured in a HAT medium to give a hybridoma. Then the hybridoma is cloned by a limiting dilution method to establish an antibody forming strain. The antibody forming strain is cultured to give the VTAT monoclonal antibody which specifically reacts with VTAT but not with liberated vitronectin, thrombin and antithrombin III.;COPYRIGHT: (C)1996,JPO