The analysis of glycosylation: a continued need for high pH anion exchangechromatography

An appreciation of the structures of the oligosaccharide chains which become attached to biomolecules (theprocess known as glycosylation), and their relevance to the biological function of the molecule concerned, has progressedrapidly in recent years with developments in site-selective protein glycosylation, oligosaccharide synthesis and in vivo targetingof oligosaccharides. These developments have necessitated the parallel development of effective analytical tools for thedetermination of the structures of glycosylation. The conclusion of studies in the 1980s and 1990s was that high pH anionexchange chromatography (HPAEC) was the most effective HPLC mode for the analysis of glycosylation. It allowed the fractionationof complex mixtures of monosaccharides or oligosaccharides, the latter in terms of charge, size, composition, anomerityand intra-chain linkages. This review reinvestigates whether HPAEC still appears to offer the most effective means ofanalysing glycosylation