Takegawa Yasuhiro, Deguchi Kisaburo, Keira Takuro, Ito Hiroki, Nakagawa Hiroaki, Nishimura Shin-Ichiro
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo 001-0021, Japan.
J Chromatogr A. 2006 Apr 28;1113(1-2):177-81. doi: 10.1016/j.chroma.2006.02.010. Epub 2006 Feb 28.
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.
同分异构寡糖和同分异构糖肽有时难以在正相（NP）柱和反相（RP）柱上分离。一种带有磺基甜菜碱基团的两性离子型亲水作用色谱柱（称为ZIC-HILIC柱）首次应用于2-氨基吡啶衍生化（PA）的人血清免疫球蛋白G（IgG）的N-聚糖和胰蛋白酶肽的分离。结果表明，ZIC-HILIC柱对同分异构N-聚糖具有很高的结构识别能力，对糖肽也具有很高的选择性。前一个特性（即结构识别）通过中性PA N-聚糖异构体的充分分离得到证明，这些异构体通常难以在NP柱和RP柱上分离。此外，值得注意的是，由同分异构N-聚糖和相同肽序列组成的IgG糖肽可以在ZIC-HILIC柱上充分分离。后一个特性（即选择性）也通过轻松分离带有/不带有N-聚糖的两个肽组得到了证明。因此，我们注意到ZIC-HILIC柱对于简单分析N-聚糖和N-糖肽样品非常有前景。
