A STRATEGY FOR ISOLATION AND STRUCTURAL ANALYSIS OF GLYCOPROTEIN OLIGOSACCHARIDES

Abstract

A general strategy is described for release, separation and structural analysis of Nand O-linked glycans from glycoproteins. Following sugar analysis, to determine the monosaccharide composition and the total carbohydrate content, the glycoprotein is degraded by chemical and/or enzymatic methods. The N- and O-linked glycans are separated and further fractionated into pure compoundsby gelfiltration, affinity chromatography and high performance ion exchange chromatography. Structural analysis is carried out by chemical analyses, fast atom bombardment mass spectrometry and 500 MHz 1H-NMR spectroscopy. Complete structural analysis is obtained of each glycan as well as determination of the glycosylationsites.