In cooperation with the Iranian Nuclear Society

Document Type : Research Paper

Authors

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.

Highlights

  1. S. Potamianos, A.D. Varvarigou, S.C. Archimandritis, “Radioimmunoscintigraphy and radioimmunotherapy in cancer: principles and application,” Anticancer Research 20, 925-948 (2000).

     

  2. P.R. Srinivas, B.S. Kramer, S. Srivatava, “Trends in biomarker resaerch for cancer detection,” Lancet Oncol, 2, 698-704 (2001).

     

  3. R. Leake, “Biological markers: Maintaining Standards,” Br. J. Cancer, 82, 1627-1628 (1997).

     

  4. A.M. Ward, J.W.F. Catto, F.C. Hamdy, “Prostate specific antigen: biology, biochmistry and available commercial assays,” Ann. Clin. Biochem., 38, 633-651 (2001).

     

  5. S. Jain, A.G. Bhojwani, J.K. Mellon, “Improving the utility of prostate apecific antigen in the diagnosis of prostate cancer: the use of PSA derivatives and novel markers,” Postgard Med, 78, 646-650 (2002).

     

  6. D. Rusciano, A. Berardi, C. Caccarini, B. Terrana, “Concomitant purification of prostate carcinoma tumor markers from human seminal fluid under nondenaturing conditions,” Clin. Chem., 34, 2528-2832 (1988).

     

  7. O.H. Lowry, N.J. Rosebrough, L. Farr, R.J. Randal, “Protein measurement with the folin phenol reagent,” J. Biol. Chem., 193, 265 (1951).

     

     

     

     

     

  8. G.C. Howard, and D.R. Bethell, “Basic methods in antibody production and characterization,” Florida, CRC Press 51-68 (2001).

     

  9. M.H. Babaei, P. Behradkia, M. Shafii, H. Forutan, R. Najafi, “Screening for pair monoclonal antibody directly on culture supernatants in the early phase of monoclonal antibody production against PSA by a RIA method,” (Submitted to J. Immunol. Methods).

     

  10. W. Zhong, L. Chen, R. Wang, “Human prostate specific antigen (hPSA) purification and establishment of hPSA radioimmunoassay,” J. Radioanal. Nucl. Chem., 206, 227-231 (1996).

     

  11. G. Kohler, and C. Milstein, “Cotinuous cultures of fused cells secreting antibod of predefined specificity,” Nature, 256, 495-497 (1975).

     

  12. M.R.A. Pillai, and S.D. Bhandarkar, “Radioimmunoassay: Principles and Practice,” Devi printers and binders, Thane, India (1998).

     

  13. R. Junker, B. Brandt, C. Zechei, G. Assmann, “Comparison of PSA measured by four combonations of free PSA and total PSA assays,” Clin. Chem., 43, 1588-1594 (1997).

Keywords

  1. S. Potamianos, A.D. Varvarigou, S.C. Archimandritis, “Radioimmunoscintigraphy and radioimmunotherapy in cancer: principles and application,” Anticancer Research 20, 925-948 (2000).

     

  2. P.R. Srinivas, B.S. Kramer, S. Srivatava, “Trends in biomarker resaerch for cancer detection,” Lancet Oncol, 2, 698-704 (2001).

     

  3. R. Leake, “Biological markers: Maintaining Standards,” Br. J. Cancer, 82, 1627-1628 (1997).

     

  4. A.M. Ward, J.W.F. Catto, F.C. Hamdy, “Prostate specific antigen: biology, biochmistry and available commercial assays,” Ann. Clin. Biochem., 38, 633-651 (2001).

     

  5. S. Jain, A.G. Bhojwani, J.K. Mellon, “Improving the utility of prostate apecific antigen in the diagnosis of prostate cancer: the use of PSA derivatives and novel markers,” Postgard Med, 78, 646-650 (2002).

     

  6. D. Rusciano, A. Berardi, C. Caccarini, B. Terrana, “Concomitant purification of prostate carcinoma tumor markers from human seminal fluid under nondenaturing conditions,” Clin. Chem., 34, 2528-2832 (1988).

     

  7. O.H. Lowry, N.J. Rosebrough, L. Farr, R.J. Randal, “Protein measurement with the folin phenol reagent,” J. Biol. Chem., 193, 265 (1951).

     

     

     

     

     

  8. G.C. Howard, and D.R. Bethell, “Basic methods in antibody production and characterization,” Florida, CRC Press 51-68 (2001).

     

  9. M.H. Babaei, P. Behradkia, M. Shafii, H. Forutan, R. Najafi, “Screening for pair monoclonal antibody directly on culture supernatants in the early phase of monoclonal antibody production against PSA by a RIA method,” (Submitted to J. Immunol. Methods).

     

  10. W. Zhong, L. Chen, R. Wang, “Human prostate specific antigen (hPSA) purification and establishment of hPSA radioimmunoassay,” J. Radioanal. Nucl. Chem., 206, 227-231 (1996).

     

  11. G. Kohler, and C. Milstein, “Cotinuous cultures of fused cells secreting antibod of predefined specificity,” Nature, 256, 495-497 (1975).

     

  12. M.R.A. Pillai, and S.D. Bhandarkar, “Radioimmunoassay: Principles and Practice,” Devi printers and binders, Thane, India (1998).

     

  13. R. Junker, B. Brandt, C. Zechei, G. Assmann, “Comparison of PSA measured by four combonations of free PSA and total PSA assays,” Clin. Chem., 43, 1588-1594 (1997).