Structural analysis of sialyl N-glycan using pyridylamination and chromatography followed by multistage tandem mass spectrometry.
Multiple-dimensional mapping (n-DM) methods consisting of pyridylamination and high-performance liquid chromatography (HPLC) are widely used for oligosaccharide analysis. These methods are quantitative, sensitive, and suitable for separating isomers. Oligosaccharide structures are suggested by elution positions on two or three kinds of columns and then confirmed by enzymatic and chemical treatments or mass spectrometry (MS). Multiple-stage tandem MS (MS(n)) analyses have been used to determine oligosaccharide structures by spectrum matching, comparing standard oligosaccharides, and offering detailed MS(n) fragment analysis. However, oligosaccharides usually exist as a mixture including isomers. The 3-DM method provides quantitative information and well-isolated sialyl- or neutral-oligosaccharides; on the other hand, the MS(n) method saves time by eliminating complicated enzyme degradation steps. The combination of both methods, which support each other, represents a reasonable and efficient strategy. This chapter describes 3-DM on HPLC after negative-ion MS(n) spectral matching for sialyl N-glycans.
Nakagawa
Hiroaki
H
Biological Sciences Laboratory of Glyco-Finechemistry, Hokkaido University, Sapporo, Japan.
Deguchi
Kisaburo
K
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis; D011134:Polysaccharides / Q000737:chemistry*; D012794:Sialic Acids / Q000737:chemistry*; D013237:Stereoisomerism
D006023:Glycoproteins; D009844:Oligosaccharides; D011134:Polysaccharides; D012794:Sialic Acids