Fractionation of plasma proteins by pseudo-ligand affinity chromatography. Elution from immobilized Brown Fractogel TSK-AF

Abstract

The fractionation of human plasma by chromatography on immobilized Brown Fractogel TSK-AF was analyzed by following the elution profile of 25 different plasma proteins. A three-step procedure was used to elute proteins from the column. First, a low-molarity buffer (30 mM sodium phosphate, pH 7.0, I = 0.053) was applied; then a linear salt gradient (0-1.0 M sodium chloride in the above buffer) was followed by an additional wash with four bed volumes of 1.0 M sodium chloride. Tightly bound proteins were finally stripped with 0.5 M ammonium thiocyanate. The elution profile of the proteins obtained by this procedure appears to be very reproducible. Comparison with the profiles obtained by chromatography on Cibacron Blue 3-GA and on Green TSK-AF indicates significant differences between the binding properties of the three gels.