Reparation And Characterization Of A Specific Monoclonal Antibody Against LAMP2Protein

Objective: The immunized BALB/c mice were prepared by preparing the immunogenicpeptide segment and anti LAMP2monoclonal antibody, and the antibody to the preliminaryidentification.Method:(1) with Boaosen LAMP2protein, and this polypeptide respectively connectedKeyhole Limpet hemocyanin (abbreviation: peptide-KLH) or bovine serum albumin (abbreviated:Peptide-BSA);(2) peptide-KLH as antigenthe immunized BALB/c mice;(3) take the serum titer isgreater than1:10000mouse spleen cells were fused with the myeloma cells SP2/0, to thepeptide-BSA package were, by enzyme-linked immunosorbent assay (ELISA) detection of hybridomasupernatants to be screened; cloned by limiting dilution of hybridoma;(4) large scale preparation ofthe monoclonal antibody was obtained by intraperitoneal injection of hybridoma cells;(5) With amouse monoclonal antibody subclass typing kit identification antibody subclasses,immunohistochemical methods showed that the preparation of monoclonal antibodies to identifyLAMP2protein.Result: A stable hybridoma cell lines secreting anti-LAMP2monoclonal antibody; usingimmunohistochemical methods to prove that the newly prepared anti-LAMP2monoclonal antibodiesrecognized LAMP2antigen; the subclasses IgG2a/κ this monoclonal antibodies.Conclusion: The obtained hybridoma cell lines stably secreting purpose monoclonal antibody;immunohistochemical methods showed that the preparation of monoclonal antibodies recognized thethe LAMP2antigen, which laid the foundation for a more in-depth study of the biological function ofthe LAMP2its subtypes, but alsothe preparation conditions for the the laboratory the late preparedLAMP2antigen ELISA kit.