Purification of prostate specific antigen and establishment of Hybridoma cell lines secreting monoclonal antibody against PSA / 中国免疫学杂志

ABSTRACT

Objective:To prepare anti-human PSA(Prostate Specific Antigen)monoclonal antibodys(McAb)and identify their distinct biological function.Methods:Prostate Specific Antigen(PSA)was purified by salting out,ion exchange and gel filtration method from human sperm.The purity and contents of the objective protein was detected by SPS-PAGE and f-PSA EiAsy.BALB/c mice were immunized with the purified PSA purified.Then the immunized splenocytes were isolated and fused with a hypoxanthine-aminopterin-thymidine-sensitive mouse myeloma cell line SP2/0.Positive clones were screened by limiting dilution method.After acquisition of the hybidomas secreting anti-PSA monoclonal antibody,investigation of their sub-class isotype,binding affinity and specificity were followed.Results:Fifteen hybridoma cell lines that could continuously secret anti-PSA monoclonal antibody were obtained.An Ig subclass was classified as IgG2a and the other were IgG1.Conclusion:This McAb is propitious to establish PSA-ELISA diagnose reagent.