An assessment of the fractionation of carbohydrates on Concanavalin A–Sepharose 4B by affinity chromatography

Abstract

Assessment of a water-insoluble but active form of the lectin Concanavalin A has established that a number of monosaccharides may be separated by elution from a column of Concanavalin A immobilised on Sepharose 4B with aqueous phosphate buffer at near neutral pH values as eluant. Branched-chain polysaccharides may also be fractionated on the column. Weakly interacting polysaccharide fractions are eluted in the phosphate buffer; more tightly bound fractions may be eluted with borate buffer. The use of borate buffer for this purpose overcomes the problems arising in the use of carbohydrates for the elution of bound polysaccharide. The work demonstrates that mixtures of carbohydrates which are not separable by complex formation with Concanavalin A in solution may be separated by use of an immobilised form of the lectin.