Separation of oligosaccharides containing terminal α-linked galactose residues by affinity chromatography on Griffonia simplicifolia I bound to concanavalin A-Sepharose

Abstract

The seeds of Griffonia simplicifolia contain a family of five isolectins (GS-I) ( L. A. Murphy and I. J. Goldstein (1977) J. Biol. Chem. 252, 4739–4742) that bind with high affinity to glycoconjugates containing terminal nonreducing α-linked galactose residues. Here, we report that GS-I itself is bound via its high mannose-type, Asn-linked sugar chains to immobilized concanavalin A (Con A-Sepharose). The GS-I in the GS-I-Con A-Sepharose complex retains its ability to bind glycoconjugates containing terminal α-linked galactose residues. This convenient method to immobilize GS-I is rapid and quantitative. We have exploited this affinity system to separate oligosaccharides based on their number of terminal α-linked d-galactose residues.