Measurement of the testosterone binding parameters for both testosterone-estradiol binding globulin and albumin in individual serum samples

Abstract

This report describes a solid phase method for the characterization of testosterone binding to both albumin and testosterone-estradiol binding globulin (TeBG). TeBG is adsorbed from serum samples onto a solid phase matrix of concanavalin A covalently linked to 4B Sepharose. The binding of testosterone is then examined both in the presence and absence of the endogenous serum albumin. Analysis of the resulting Scatchard plots permits determination of the TeBG binding capacity, TeBG association constant and a parameter of albumin binding equivalent to the product of its affinity and capacity for binding testosterone. Results showed that the TeBG capacity was lower in men than in women (10.4 ± 5.8 vs. 33.1 ± 19.2 nM, p < 0.01). The association constant was greater in men (1.59 ± 0.35 vs. 1.19 ± 0.32 × 10 9M −1, 10 9M −1. p < 0.01). There was no difference in the albumin binding parameter (43.8 ± 18.3 vs. 46.6 ± 15.5, NS). These parameters can then be used to calculate the distribution of the circulating testosterone into albumin bound, TeBG bound and unbound fractions.