Development of a New in Vivo Kit for Detection of Prostate Specific Antigen in Human Serum Using Immunoradiometric Assay Method
M. H. Babaei, Behrad Kia Payam, S. Fazal Mohammad, M. M. Moula, Haleh Foroutan, Najafi Reza
Abstract
M. H. Babaei, Behrad Kia Payam, S. Fazal Mohammad, M. M. Moula, Haleh Foroutan, Najafi Reza
Abstract
Prostate is a leading site for the cancer incidence, accounted for 31.0% of new cancer cases in men. Prostate-specific antigen (PSA) is widely used in the detection and monitoring of the prostate cancer. Currently, immunoassay is used to detect PSA in human serum. This technique is based on the interaction between antibody and antigen. The varied immunoassay formats and equipment to run the assays allow the users to measure the analytes rapidly, with the flexibility to run a small or a large number of samples. Among different immunoassay methods, immunoradiometric assay (IRMA) is a more sensitive and valuable detection approach. This study has been made in 4 parts: (1) purification of PSA from seminal fluid; (2) preparation of hybridoma cells which secrete monoclonal antibody (mAb) against PSA, (3) selection of pair monoclonal antibody among those antibodies, and finally (4) design of an IRMA kit and it's quality control . The results of this study were: (1) obtaing a huge amount of PSA as semi-purified and purified, that is a valuable material for preparation of standard kits; (2) preparation of 8 kinds of monoclonal antibodies; (3) finding 4 pairs of monoclonal antibodies which react with different epitopes on PSA molecule; and (4) preparation of IRMA kit for measuring PSA concentration in human serum.
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Prostate is a leading site for the cancer incidence, accounted for 31.0% of new cancer cases in men. Prostate-specific antigen (PSA) is widely used in the detection and monitoring of the prostate cancer. Currently, immunoassay is used to detect PSA in human serum. This technique is based on the interaction between antibody and antigen. The varied immunoassay formats and equipment to run the assays allow the users to measure the analytes rapidly, with the flexibility to run a small or a large number of samples. Among different immunoassay methods, immunoradiometric assay (IRMA) is a more sensitive and valuable detection approach. This study has been made in 4 parts: (1) purification of PSA from seminal fluid; (2) preparation of hybridoma cells which secrete monoclonal antibody (mAb) against PSA, (3) selection of pair monoclonal antibody among those antibodies, and finally (4) design of an IRMA kit and it's quality control . The results of this study were: (1) obtaing a huge amount of PSA as semi-purified and purified, that is a valuable material for preparation of standard kits; (2) preparation of 8 kinds of monoclonal antibodies; (3) finding 4 pairs of monoclonal antibodies which react with different epitopes on PSA molecule; and (4) preparation of IRMA kit for measuring PSA concentration in human serum.
Key concepts: Immunoradiometric assay, Immunoassay, Monoclonal antibody, Antigen, Prostate-specific antigen, Epitope, Prostate cancer, Radioimmunoassay