MONOCLONAL ANTIBODY AGAINST CRK PROTEIN AND HYBRIDOMA STRAIN PRODUCING THE ANTIBODY

摘要

PURPOSE:To obtain a new high-precision reagent having specific reactivity to CRK protein which is an expression product of precancer gene CRK and useful for the basic research of human cancer cell and to obtain the subject monoclonal antibody useful for the development of new anticancer agent. CONSTITUTION:An E.coli is transformed with an expression vector produced by connecting and inserting human CRK gene to the 3' terminal of glutathione-S- transferase gene. CRK protein purified from the transformed E.coli by affinity chromatography is administered to a mouse in several divided doses with intervals of several weeks and the lymphocyte is collected from the spleen after the final immunization. The lymphocyte is fused with a myeloma cell originated from mouse and cultured by selective culture in HAT medium to obtain hybridoma, an antibody-producing hybridoma is selected therefrom, the selected hybridoma is monoclonized by cloning e.g. using a limiting dilution culture and the obtained hybridoma AD2 strain (FERM P-13176) is cultured in a medium to obtain the objective monoclonal antibody exhibiting specific reactivity with CRK protein.