Characterization and measurement of prostate-specific antigen using monoclonal antibodies.

Abstract

Monoclonal and polyclonal antibodies were produced using a pure preparation of prostate-specific antigen (PSA) as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). By SDS-PAGE, the apparent molecular weight of PSA was about 33 kD. Eighty-four monoclonal antibodies were produced, 77 of which bind PSA with an affinity higher than 10(9) M-1. Use of these monoclonal antibodies to study the immunological characteristics of PSA revealed the presence of 4 epitopes. Injection of PSA into goats resulted in a production of polyclonal antibodies with high affinity. These polyclonal antibodies were purified by affinity chromatography and adsorbed on plastic tubes. By an immunometric assay, we have also demonstrated that polyclonal antibodies bind PSA at a fifth epitope that is different from those of monoclonal antibodies. Using an iodinated monoclonal antibody and polyclonal antibodies adsorbed on plastic tubes, a sensitive immunoradiometric assay could be developed, and a further increase in sensitivity could be achieved by using a mixture of 2 monoclonal antibodies. The serum PSA levels in 2,250 patients measured with this immunoradiometric assay were identical to the values determined by Tandem-R, although the present assay reached a minimum detectable value of 0.05 ng/ml compared with 0.2 ng/ml by Tandem-R.