Preparation And Application Of Functional Monoclonal Antibody Against Human B7-2 Molecule

ObjectiveTo obtain a hybridoma cell line that can stably secret mouse anti-human B7-2 monoclonal antibody and study its biological characteristics. To establish a sensitive, specific, stable ELISA assay for detecting soluble human B7-2 molecule.MethodsB lymphocyte hybridoma technique was applied to obtain a hybridoma cell line secreting anti-human B7-2 antibody. The subclass of the mAb was identified through fast-strip method analysis. Ascites were induced to produce the monoclonal antibody from BALB/c mouse, and were purified with affinity chromatography method. Indirect immunofluorescence was used to test the abiliby of mAb to recognize the B7-2 molecule on different cell membranes. MTT assay was used to detect its biological effects of inhibiting Daudi and PBTCs proliferation. The epitope recognition was identified through antigen competitive inhibition assays. Antibodies was labeled with biotin to establish the ELISA method for measuring soluble B7-2.ResultsObtained a hybridoma named 6A5 which can stably secreting monoclonal antibody against human B7-2 molecule. The subclass of 6A5 was mouse IgG1 with kappa light chains through fast-strip method analysis. Ascites were induced to produce the monoclonal antibody and 5.9 ml ascites was obtained from each BALB/c mouse on...