title: Separation of isomeric 2-aminopyridine derivatized N-glycans and N-glycopeptides of human serum immunoglobulin G by using a zwitterionic type of hydrophilic-interaction chromatography.
abstract: Isomeric oligosaccharides and isomeric glycopeptides are sometimes difficult to separate on normal-phase (NP) and reversed-phase (RP) columns. A zwitterionic type of hydrophilic-interaction chromatography column with sulfobetaine groups (called ZIC-HILIC column) was first applied to the separation of 2-aminopyridine derivatized (PA) N-glycans and tryptic peptides of human serum immunoglobulin G (IgG). It is shown that the ZIC-HILIC column has high capability for structural recognition of isomeric N-glycans as well as high selectivity for glycopeptides. The former feature (i.e., structural recognition) was proven by sufficient separation of neutral PA N-glycan isomers, which are usually difficult to separate on NP and RP columns. In addition, it is noteworthy that IgG glycopeptides consisting of isomeric N-glycans and the same peptide sequences can be sufficiently separated on a ZIC-HILIC column. The latter feature (i.e., selectivity) was also demonstrated by easily separating two peptide groups with/without N-glycans. Thus, we note that the ZIC-HILIC column is highly promising for a simple analysis of N-glycans and N-glycopeptide samples.
authors.affiliation: Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takegawa
authors.forename: Kisaburo
authors.initials: K
authors.lastname: Deguchi
authors.forename: Takuro
authors.initials: T
authors.lastname: Keira
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ito
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nakagawa
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: D000631:Aminopyridines / Q000737:chemistry / Q000302:isolation & purification*; D002853:Chromatography, Liquid / Q000379:methods*; D006020:Glycopeptides / Q000737:chemistry*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood*; D007536:Isomerism; D011134:Polysaccharides / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D000631:Aminopyridines; D006020:Glycopeptides; D007074:Immunoglobulin G; D011134:Polysaccharides; C032439:alpha-aminopyridine