Resolution of nucleotide sugars and oligosaccharides by lectin affinity chromatography

Abstract

Bandeiraea simplicifolia I (BS I) isolectins, immobilized on Sepharose beads, specifically retarded low molecular weight ligands containing terminal [alpha]--galactopyranosyl and 2-acetamido-2-deoxy-[alpha]--galactopyranosyl residues. A BS I lectin-Sepharose column has been used to perform very efficient separations of structurally homologous sugar nucleotides and oligosaccharides. For example, UDP-glucose and UDP-galactose (and the corresponding 2-acetamido-2-deoxy derivatives) were separated by as much as three column volumes by a BS I lectin-Sepharose column. This column has also been used to resolve the alditols of lactose ([beta]--Galp-(1 --> 4)--Glc) and melibiose ([alpha]--Galp-(1 --> 6)--Glc), to analyze the radiochemical purity of UDP-galactose and UDP-N-acetyl--galactosamine preparations, and to analyze the relative proportions of UDP-N-acetyl--glucosamine and UDP-N-acetyl--galactosamine in the UDP-N-acetyl--hexosamine pool isolated from cultured cells.