Skip to main content
British Journal of Cancer logoLink to British Journal of Cancer
. 1997;75(6):789–797. doi: 10.1038/bjc.1997.142

Antigenic determinants of prostate-specific antigen (PSA) and development of assays specific for different forms of PSA.

O Nilsson 1, A Peter 1, I Andersson 1, K Nilsson 1, B Grundström 1, B Karlsson 1
PMCID: PMC2063392  PMID: 9062397

Abstract

Monoclonal antibodies were raised against prostate-specific antigen (PSA) by immunization with purified free PSA, i.e. not in complex with any protease inhibitor (F-PSA) and PSA in complex with alpha1-anti-chymotrypsin (PSA-ACT). Epitope mapping of PSA using the established monoclonal antibody revealed a complex pattern of independent and partly overlapping antigenic domains in the PSA molecule. Four independent antigenic domains and at least three partly overlapping domains were exposed both in F-PSA and in the PSA-ACT complex, while one antigenic domain was specific for F-PSA. The different domains contained both continuous and discontinuous epitopes. The combination of antibodies recognizing antigenic domains exposed both in F-PSA and PSA-ACT made it possible to develop several highly sensitive sandwich immunoassays for determination of total PSA, i.e. F-PSA + PSA-ACT, with the same molar response for F-PSA and PSA-ACT. Assays specific for F-PSA (cross-reactivity between F-PSA and PSA-ACT < 1%) were developed by the combination of antibodies recognizing epitopes exposed only in F-PSA and antibodies recognizing epitopes exposed both in F-PSA and PSA-ACT.

Full text

PDF
789

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akiyama K., Nakamura T., Iwanaga S., Hara M. The chymotrypsin-like activity of human prostate-specific antigen, gamma-seminoprotein. FEBS Lett. 1987 Dec 10;225(1-2):168–172. doi: 10.1016/0014-5793(87)81151-1. [DOI] [PubMed] [Google Scholar]
  2. Bélanger A., Rong P. M., Liu S., Lavoie L., Labrie F. Characterization and measurement of prostate-specific antigen using monoclonal antibodies. Clin Invest Med. 1993 Dec;16(6):409–414. [PubMed] [Google Scholar]
  3. Christensson A., Björk T., Nilsson O., Dahlén U., Matikainen M. T., Cockett A. T., Abrahamsson P. A., Lilja H. Serum prostate specific antigen complexed to alpha 1-antichymotrypsin as an indicator of prostate cancer. J Urol. 1993 Jul;150(1):100–105. doi: 10.1016/s0022-5347(17)35408-3. [DOI] [PubMed] [Google Scholar]
  4. Christensson A., Laurell C. B., Lilja H. Enzymatic activity of prostate-specific antigen and its reactions with extracellular serine proteinase inhibitors. Eur J Biochem. 1990 Dec 27;194(3):755–763. doi: 10.1111/j.1432-1033.1990.tb19466.x. [DOI] [PubMed] [Google Scholar]
  5. Chu T. M., Kawinski E., Hibi N., Croghan G., Wiley J., Killian C. S., Corral D. Prostate-specific antigenic domain of human prostate specific antigen identified with monoclonal antibodies. J Urol. 1989 Jan;141(1):152–156. doi: 10.1016/s0022-5347(17)40630-6. [DOI] [PubMed] [Google Scholar]
  6. Graves H. C. Standardization of immunoassays for prostate-specific antigen. A problem of prostate-specific antigen complexation or a problem of assay design? Cancer. 1993 Dec 1;72(11):3141–3144. doi: 10.1002/1097-0142(19931201)72:11<3141::aid-cncr2820721104>3.0.co;2-1. [DOI] [PubMed] [Google Scholar]
  7. Graves H. C., Wehner N., Stamey T. A. Comparison of a polyclonal and monoclonal immunoassay for PSA: need for an international antigen standard. J Urol. 1990 Dec;144(6):1516–1522. doi: 10.1016/s0022-5347(17)39789-6. [DOI] [PubMed] [Google Scholar]
  8. Hemmilä I., Dakubu S., Mukkala V. M., Siitari H., Lövgren T. Europium as a label in time-resolved immunofluorometric assays. Anal Biochem. 1984 Mar;137(2):335–343. doi: 10.1016/0003-2697(84)90095-2. [DOI] [PubMed] [Google Scholar]
  9. Leinonen J., Lövgren T., Vornanen T., Stenman U. H. Double-label time-resolved immunofluorometric assay of prostate-specific antigen and of its complex with alpha 1-antichymotrypsin. Clin Chem. 1993 Oct;39(10):2098–2103. [PubMed] [Google Scholar]
  10. Lilja H. A kallikrein-like serine protease in prostatic fluid cleaves the predominant seminal vesicle protein. J Clin Invest. 1985 Nov;76(5):1899–1903. doi: 10.1172/JCI112185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lilja H., Christensson A., Dahlén U., Matikainen M. T., Nilsson O., Pettersson K., Lövgren T. Prostate-specific antigen in serum occurs predominantly in complex with alpha 1-antichymotrypsin. Clin Chem. 1991 Sep;37(9):1618–1625. [PubMed] [Google Scholar]
  12. Lilja H., Cockett A. T., Abrahamsson P. A. Prostate specific antigen predominantly forms a complex with alpha 1-antichymotrypsin in blood. Implications for procedures to measure prostate specific antigen in serum. Cancer. 1992 Jul 1;70(1 Suppl):230–234. doi: 10.1002/1097-0142(19920701)70:1+<230::aid-cncr2820701310>3.0.co;2-y. [DOI] [PubMed] [Google Scholar]
  13. Lilja H., Oldbring J., Rannevik G., Laurell C. B. Seminal vesicle-secreted proteins and their reactions during gelation and liquefaction of human semen. J Clin Invest. 1987 Aug;80(2):281–285. doi: 10.1172/JCI113070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lindholm L., Holmgren J., Svennerholm L., Fredman P., Nilsson O., Persson B., Myrvold H., Lagergård T. Monoclonal antibodies against gastrointestinal tumour-associated antigens isolated as monosialogangliosides. Int Arch Allergy Appl Immunol. 1983;71(2):178–181. doi: 10.1159/000233384. [DOI] [PubMed] [Google Scholar]
  15. Lundwall A. Characterization of the gene for prostate-specific antigen, a human glandular kallikrein. Biochem Biophys Res Commun. 1989 Jun 30;161(3):1151–1159. doi: 10.1016/0006-291x(89)91362-4. [DOI] [PubMed] [Google Scholar]
  16. Lundwall A., Lilja H. Molecular cloning of human prostate specific antigen cDNA. FEBS Lett. 1987 Apr 20;214(2):317–322. doi: 10.1016/0014-5793(87)80078-9. [DOI] [PubMed] [Google Scholar]
  17. Lövgren J., Piironen T., Overmo C., Dowell B., Karp M., Pettersson K., Lilja H., Lundwall A. Production of recombinant PSA and HK2 and analysis of their immunologic cross-reactivity. Biochem Biophys Res Commun. 1995 Aug 24;213(3):888–895. doi: 10.1006/bbrc.1995.2212. [DOI] [PubMed] [Google Scholar]
  18. Oesterling J. E. Prostate specific antigen: a critical assessment of the most useful tumor marker for adenocarcinoma of the prostate. J Urol. 1991 May;145(5):907–923. doi: 10.1016/s0022-5347(17)38491-4. [DOI] [PubMed] [Google Scholar]
  19. Pettersson K., Piironen T., Seppälä M., Liukkonen L., Christensson A., Matikainen M. T., Suonpä M., Lövgren T., Lilja H. Free and complexed prostate-specific antigen (PSA): in vitro stability, epitope map, and development of immunofluorometric assays for specific and sensitive detection of free PSA and PSA-alpha 1-antichymotrypsin complex. Clin Chem. 1995 Oct;41(10):1480–1488. [PubMed] [Google Scholar]
  20. Prestigiacomo A. F., Stamey T. A. A comparison of 4 ultrasensitive prostate specific antigen assays for early detection of residual cancer after radical prostatectomy. J Urol. 1994 Nov;152(5 Pt 1):1515–1519. doi: 10.1016/s0022-5347(17)32459-x. [DOI] [PubMed] [Google Scholar]
  21. Prestigiacomo A. F., Stamey T. A. Clinical usefulness of free and complexed PSA. Scand J Clin Lab Invest Suppl. 1995;221:32–34. doi: 10.3109/00365519509090561. [DOI] [PubMed] [Google Scholar]
  22. Riegman P. H., Vlietstra R. J., van der Korput J. A., Romijn J. C., Trapman J. Characterization of the prostate-specific antigen gene: a novel human kallikrein-like gene. Biochem Biophys Res Commun. 1989 Feb 28;159(1):95–102. doi: 10.1016/0006-291x(89)92409-1. [DOI] [PubMed] [Google Scholar]
  23. Schedlich L. J., Bennetts B. H., Morris B. J. Primary structure of a human glandular kallikrein gene. DNA. 1987 Oct;6(5):429–437. doi: 10.1089/dna.1987.6.429. [DOI] [PubMed] [Google Scholar]
  24. Stamey T. A., Prestigiacomo A. F., Chen Z. Standardization of immunoassays for prostate specific antigen. A different view based on experimental observations. Cancer. 1994 Sep 15;74(6):1662–1666. doi: 10.1002/1097-0142(19940915)74:6<1662::aid-cncr2820740604>3.0.co;2-k. [DOI] [PubMed] [Google Scholar]
  25. Vessella R. L., Lange P. H. Issues in the assessment of PSA immunoassays. Urol Clin North Am. 1993 Nov;20(4):607–619. [PubMed] [Google Scholar]
  26. Vessella R. L. Trends in immunoassays of prostate-specific antigen: serum complexes and ultrasensitivity. Clin Chem. 1993 Oct;39(10):2035–2039. [PubMed] [Google Scholar]
  27. Wang M. C., Valenzuela L. A., Murphy G. P., Chu T. M. Purification of a human prostate specific antigen. Invest Urol. 1979 Sep;17(2):159–163. [PubMed] [Google Scholar]
  28. Watt K. W., Lee P. J., M'Timkulu T., Chan W. P., Loor R. Human prostate-specific antigen: structural and functional similarity with serine proteases. Proc Natl Acad Sci U S A. 1986 May;83(10):3166–3170. doi: 10.1073/pnas.83.10.3166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Wood W. G., van der Sloot E., Böhle A. The establishment and evaluation of luminescent-labelled immunometric assays for prostate-specific antigen-alpha 1-antichymotrypsin complexes in serum. Eur J Clin Chem Clin Biochem. 1991 Dec;29(12):787–794. doi: 10.1515/cclm.1991.29.12.787. [DOI] [PubMed] [Google Scholar]
  30. Yu H., Diamandis E. P. Ultrasensitive time-resolved immunofluorometric assay of prostate-specific antigen in serum and preliminary clinical studies. Clin Chem. 1993 Oct;39(10):2108–2114. [PubMed] [Google Scholar]
  31. Zhou A. M., Tewari P. C., Bluestein B. I., Caldwell G. W., Larsen F. L. Multiple forms of prostate-specific antigen in serum: differences in immunorecognition by monoclonal and polyclonal assays. Clin Chem. 1993 Dec;39(12):2483–2491. [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES