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

Journal of Chromatography. a
Yasuhiro TakegawaShin-Ichiro Nishimura

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.

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Citations

Aug 14, 2012·Glycoconjugate Journal·Jonas NilssonGöran Larson
Sep 5, 2012·Glycoconjugate Journal·Milos V NovotnyBenjamin F Mann
Jun 29, 2013·Journal of Chromatography. a·Zhaorui ZhangMary J Wirth
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Mar 9, 2011·Journal of Pharmaceutical and Biomedical Analysis·Keita Yamada, Kazuaki Kakehi
Dec 15, 2010·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·David J Harvey
Dec 29, 2009·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Joomi AhnMartin Gilar
Feb 26, 2008·Journal of Chromatography. a·Tohru IkegamiNobuo Tanaka
Aug 15, 2014·Fungal Genetics and Biology : FG & B·Yi QuSi Wu
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Sep 22, 2012·Journal of Separation Science·Sara OngayRainer Bischoff
Oct 24, 2012·Proteomics. Clinical Applications·Shuang Yang, Hui Zhang

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