Preparation And Characterization Of Anti-prostate Stem Cell Antigen (PSCA) Monoclonal Antibody

Part I The preparation of anti-prostate stem cell antigen monoclonal antibodyPURPOSES: To prepare the hybridoma producing specific monoclonal antibody against prostate stem cell antigen .METHODS: 1) The spleen cells of the mice immunized with PC-3 prostate cancer cells were fused with SP2/0 myeloma cells by PEG and cultured with HAT ; 2) By ELISA screening, to check out positive clones ; 3) By muti-subcloning to promise all the hybridoma cells secret monoclonal;4) By injecting hybridoma cells into the abdomen of mice, acquire ascites containing monoclonal antibody.RESULTS: 1, Hybridoma cells grew in 176 wells ; 2,12 wells were checked out as positive ; 3, All hybridoma cells were testified to secret monoclonal antibody ; 4, A large amount of ascites was acquired.CONCLUSIONS: One hybridoma secreting specific monoclonal antibody against PC-3 prostate cancer cell is preliminarily established.Part II Functional and biochemical characterization of monoclonal antibodyPURPOSE: To characterize the monoclonal antibody produce by hybridoma for further application .METHOD: 1. To make sure the subclass of the monoclonalantibody by double-way immunodifrusion; 2. Assess the monoclonal antibody's immunoreaction with PC-3 cells ; 3. The reaction spectrum of the monoclonal antibody was checked by immunohistochemical staining in tissue sections.RESULT: 1.An immunoprecipitation line appeared between monoclonal antibody and anti-IgGl;2.The monoclonal antibody demonstrated positive reaction with prostate cancer , but negative reaction with non-prostate cancer tumor samples. As far as normal human tissue is concern, no positive reaction happened.CONCLUSION: The monoclonal antibody belongs to IgGl with high specificity and quality.