A new principle in biospecific affinity chromatography used for purification of cobalamin-binding proteins

Abstract

To avoid using protein-denaturing agents for desorption, when purifying cobalamin-binding protein by biospecific affinity chromatography, an affinity column has been prepared where cobalamin is attached through a temperature-labile linkage to insolubilized 3,3′-diaminodipropylamine. On desorption the protein is obtained in solution saturated with cobalamin. The method has been used for purification of intrinsic factor and transcobalamin I.