Enzyme immunoassay of the glycoprotein tropic hormones--choriogonadotropin, lutropin, thyrotropin--with solid-phase monoclonal antibody for the alpha-subunit and enzyme-coupled monoclonal antibody specific for the beta-subunit.

Abstract

Abstract Monoclonal antibody technology has made it possible to produce homogeneous populations of antibodies to discrete determinants on an antigen surface. We have produced monoclonal antibodies to the alpha-subunit and beta-subunits of the glycoprotein hormones choriogonadotropin, thyrotropin, and lutropin, and developed two-site simultaneous enzyme-linked immunospecific assays for these hormones. The anti-alpha-subunit monoclonal antibody was used as the solid-phase (coated tube) capture antibody for all three hormones; the anti-beta-subunit monoclonal antibodies were coupled to horseradish peroxidase (EC 1.11.1.7). Cross reactions between the closely related choriogonadotropin and lutropin were apparently greater in this method than in RIA, with use of the same antibodies. Ka of the antibodies did not appear to be as critical to sensitivity of the sandwich assay as it was for RIA. The lower limit of detection was 0.2 microgram/L after a 2-h incubation with serum sample at room temperature and a 30-min incubation with enzyme substrate at room temperature after washing away excess enzyme conjugate. Within-assay precision (CV) was very good, less than 6%.