Nature of cross-reaction between hCG and anti-oLH serum and development of a radioimmunoassay to measure hLH specifically in the presence of hCG

Abstract

Immunological cross-reaction between hCG and anti-oLH sera has been demonstrated using radioimmunoassay techniques. The results indicate that this cross-reaction is incomplete and that the anti-oLH sera used have the ability to distinguish between LH and hCG. Following absorption with purified hCG, anti-oLH serum was used to develop a heterologous radioimmunoassay "[125I]iodo-hLH + anti-oLH serum" (H-O, RIA) which specifically and selectively measures hLH in serum samples containing both hLH and hCG. In this radioimmunoassay hCG and subunits of hCG do not cross-react with hLH, in the range in which these hormones are present in human serum under physiological conditions. Other hormones such as hPL, hPRL, hGH, hFSH, hTSH, and GnRH do not interfere with the measurement of LH by radioimmunoassay. The sensitivity of the assay was 1.5 mIU (25 ng) per ml (LER 907 standard), and the inter- and intra-assay coefficients of variations for samples were 10.83% and 8.4%, respectively. The recoveries of hLH added to pregnancy serum containing an hCG concentration of 8.55 IU/ml were in the range 95-108%. Determination of LH content of human pituitary extracts by H-O RIA gave values which were in close agreement with those derived by bioassay (indices of discrimination 0.72-1.12). Serum LH patterns in women during normal menstrual cycles as well as in amenorrheic patients who received GnRH treatment are comparable to those reported by other investigators using other radioimmunoassay systems. Serum samples obtained during the first trimester of pregnancy, when analyzed by H-O RIA, showed basal LH levels.