Preparation and identification of monoclonal antibodies against human cysteine and glycine-rich protein 2

Objective To obtain monoclonal antibodies against cysteine and glycine-rich protein 2(CRP2) for further study of the biological function of CRP2 protein.Methods Balb/c mice were immunized with recombinant CRP2. Hybridoma cell lines secreting monoclonal antibodies against CRP2 were screened by regular cell fusion and subcloning approach.The specificities of these monoclonal antibodies were determined by ELISA,Western blotting and immunoprecipitation.Results Two hybridoma cell lines(F001 and F002) stable in secreting specific monoclonal antibodies were successfully obtained.They specifically bind to recombinant CRP2 and endogenous CRP2 proteins proved by ELISA,Western blotting.In addition,these two monoclonal antibodies could be precipitated the endogenous CRP2 protein by immunoprecipitation assay.Conclusion Monoclonal antibodies against CRP2 with high titers and specificity have been successfully prepared,providing foundation for further study of CRP2 protein.