Study of steroid-protein binding by a novel “two-tier” column employing cibacron blue F3G-A-Sepharose 4B. I—Sex hormone binding globulin

Cibracron Blue F3G-A-Sepharose gel was utilized as an affinity matrix for the removal of albumin from plasma prior to gel filtration on Sephadex LH-20.

Abstract

Cibracron Blue F3G-A-Sepharose gel was utilized as an affinity matrix for the removal of albumin from plasma prior to gel filtration on Sephadex LH-20. The incorporation of affinity chromatography and gel filtration in the same column allows for direct and simultaneous evaluation of steroid binding to both non-specific and specific binding components in human plasma. The binding parameters of Sex Hormone Binding Globulin, SHBG, are presented. The application of 5α-Dihydrotestosterone-3(O-carboxymethyl) oxime-human serum albumin, DHT-3-CMO-HSA, conjugate used in conjunction with Cibacron Blue F3G-A-Sepharose is discussed together with results on the relative affinity of steroid derivatives pertinent to the affinity purification of SHBG. The usefulness of this affinity matrix in enrichment of steroid haptens and its possible application to purifying antibodies is also discussed.