Differences in serum luteinizing hormone measurements by Immunoradiometric assay induced by kinetic manipulation of assay conditions are dependent on the endocrine milieu of serum

Divergent estimates for luteinizing hormone (LH) in individual serum samples may be given by different immunoassays. In order to investigate this phenomenom further, we have studied the effect of differences in assay kinetics within the same immunoradiometric assay (IRMA) configuration on LH measurement in sera from different endocrine states. Three pairs of monoclonal/polyclonal two-site IRMA systems for LH were developed from three LH monoclonal antibodies and a common polyclonal antihuman chorionic gonadotrophin. For IRMA systems a short and long assay, which were different only with respect to the incubation time (1/2h and overnight respectively), of the labelled monoclonal first antibody were performed. The IRMAs were all standardized against the LH international reference preparation 68/40. LH concentrations were measured by all the IRMAs in sera obtained from normal men (n = 11) and from women with polycystic ovarian syndrome (PCO; n = 13). In normal men, there were no differences in LH estimates between the short and the long assays of the three IRMA systems, and the ratios of long to short assays were similar for all the systems. However, in PCO there were significant differences between short and long assays and the ratios of long to short assays were different for the IRMA systems. These results indicate that kinetic differences between IRMAs of the same antibody configuration can be associated with differences in measured LH concentrations, depending on the endocrine status of the sera studied. As LH glycoform patterns are known to differ between normal men and PCO, the observed changes in LH estimates may be due to the different glycoform composition.