[Preparation and characterization of monoclonal antibodies for human BCSC-1 protein].

AIM: To obtain monoclonal antibodies against human BCSC-1 protein for further study of the structure and function of human BCSC-1 protein.METHODS: pET-30a-BCSC-1 plasmid was constructed and transformed into E.coli BL21(DE3) to express recombinant protein.BALB/c mice were immunized with recombinant human BCSC-1 protein,hybridoma cell lines secreting monoclonal antibodies against human BCSC-1protein were screened by regular cell fusion and subcloning approach.The specificity of these monoclonal antibodies were determined by ELISA and Western blotting.Expression of BCSC-1 protein in pcDNA3.1/v5-HisB-BCSC-1transfected MCF-7 cells was identified by Immunohistochemistry.RESULTS: Successfully constructed a prokaryotic expression vector pET30a-BCSC-1.BCSC-1 protein was expressed in E.coli BL21(DE3).One hybridoma cell line 1B3 stable in secreting specific monoclonal antibodies wa successfully obtained.It could bind specifically to BCSC-1 protein proved by Western blotting and Immunohistochemistry assay.CONCLUSION: Monoclonal antibodies of high specificity against human BCSC-1 protein has been successfully prepared,which laid the foundation for the further study of human BCSC-1 protein.