STRUCTURAL ANALYSIS OF THE CARBOHYDRATE MOIETIES OF GLYCOPROTEINS BY REGIOSPECIFIC DEGRADATION AND LIQUID CHROMATOGRAPHY

Article abstract of DOI:10.1016/0008-6215(83)88043-4

The reaction sequence of hydrazinolysis, nitrosation, and reduction, followed by liquid chromatography (l.c.) has been studied as a method for the routine structural analysis of theasparagine-bound oligosaccharides of glycoproteins.Glycopeptides derived from IgM and ovalbumin by proteolysis were used as test materials.The hydrazinium sulphate-catalysed hydrazinolysis was superior to the longer uncatalysed reaction, in that there was less non-specific degradation and a higher degree of N-deacetylation.The nitrosation products were reduced in situ with sodium cyanoborohydride, and the l.c. analysis required 20 min for the fractionation of oligosaccharides up to decasaccharide.The l.c. profile is characteristic of the structure of the carbohydrate unit. the analytical l.c. column may also be used to isolate oligosaccharide fractions in quantities of several hundred micrograms.