7] Resolution of carbohydrates by lectin affinity chromatography

Abstract

Immobilized lectins have been used successfully for many years to isolate glycoconjugates with specific carbohydrate structures. Most such applications have involved binding the glycoconjugate to a lectin-Sepharose column and subsequent elution of bound material with ligands of low molecular weight. Only in recent years have workers in a number of laboratories begun to realize the potential of lectin columns for the recognition of subtle differences in oligosaccharide structures. This chapter describes the use of a lectin column (Bandeiraea simplicifolia I (BS I)-Sepharose) that recognizes and interacts with terminal a-linked D-galactosyl (D-Galp) and N-acetyl-t)-galactosaminyl (Gal- NAcp) residues. The column can be used to resolve mixtures of nucleotide sugars and linear oligosaccharides. The separations are based on the ability of the lectin column to retard rather than bind the interacting saccharides. It is the intent, moreover, to communicate the essence of technique that may be applied to a wide range of analyses.