Simple separation of isomeric sialylated N-glycopeptides by a zwitterionic type of hydrophilic interaction chromatography
Simple separation of isomeric sialylated N-glycopeptides by a zwitterionic type of hydrophilic interaction chromatography
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DOI：
10.1002/jssc.200600133
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发表时间：
2006-11-01
影响因子：
3.1
通讯作者：
Nishimura, Shin-Ichiro
中科院分区：
文献类型：
--
作者：
Takegawa, Yasuhiro;Deguchi, Kisaburo;Nishimura, Shin-Ichiro
天冬酰胺连接的寡糖（n -聚糖）通常具有结构异质性，特别是在含有唾液化n -聚糖的蛋白质中，因此其结构分析仍然非常困难。采用带磺基甜菜碱官能团的两性离子型亲水相互作用色谱柱（ZIC-HILIC柱）分离α -l-酸糖蛋白的色氨酸肽。结果表明，ZIC-HILIC分离柱对唾液化n -糖肽具有选择性，并且基于对唾液化n -糖异构体以及先前报道的中性n -聚糖和n -糖肽的结构识别，具有很高的分离能力。2-氨基吡啶衍生的中性和唾液化n -聚糖（PA n -聚糖）的保留特性表明，n -聚糖的保留主要基于与ZIC-HILIC柱表面生成的富水液层的亲水性相互作用。此外，反离子屏蔽的静电斥力相互作用有效地调节了唾液化n -聚糖异构体的分离和识别。
Asparagine-linked oligosaccharides (N-glycans) usually show structural heterogeneity, especially in proteins with sialylated N-glycans and, therefore, their structural analysis is still very difficult. A zwitterionic type of hydrophilic interaction chromatography column with sulfobetaine functional groups (called a ZIC-HILIC column) was applied to the separation of tryptic peptides of alpha-l-acid glycoprotein. It was demonstrated that the ZIC-HILIC separation column has a selectivity for sialylated N-glycopeptides and a high capability for separation based on the structural recognition of sialylated N-glycan isomers as well as for the previously reported neutral N-glycans and N-glycopeptides. The retention characteristics of neutral and sialylated N-glycans derivatized with 2-aminopyridine (PA N-glycans) demonstrate that the retentions of the N-glycans are based primarily on hydrophilic interaction with the water-rich liquid layer generated on the surface of the ZIC-HILIC column. In addition, the electrostatic repulsion interaction shielded with counter ions effectively tunes the separation and recognition of sialylated N-glycan isomers.