Separation of 2-aminobenzamide labeled glycans using hydrophilic interaction chromatography columns packed with 1.7 μm sorbent

被引：160

作者：

Ahn, Joomi ^{[1
]}

Bones, Jonathan ^{[2
]}

Yu, Ying Qing ^{[1
]}

Rudd, Pauline M. ^{[2
]}

Gilar, Martin ^{[1
]}

机构：

[1] Waters Corp, Biopharmaceut Sci, Milford, MA 01757 USA

[2] Univ Coll Dublin, Dublin Oxford Glycobiol Lab, NIBRT, Dublin 4, Ireland

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2010年 / 878卷 / 3-4期

关键词：

Glycans; Oligosaccharides; Ultra-performance liquid chromatography; Hydrophilic interaction chromatography; 2-Aminobenzamide labeled glycans; PERFORMANCE LIQUID-CHROMATOGRAPHY; ANION-EXCHANGE CHROMATOGRAPHY; GRAPHITIZED CARBON COLUMN; N-LINKED OLIGOSACCHARIDES; MASS-SPECTROMETRY; HIGH-RESOLUTION; PEAK-CAPACITY; GLYCOPEPTIDES; OLIGONUCLEOTIDES; GLYCOSYLATION;

D O I：

10.1016/j.jchromb.2009.12.013

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Separation by hydrophilic interaction chromatography (HILIC) with fluorescence detection utilizing a sub-2 mu m glycan column for the separation of 2-aminobenzamide (2-AB) labeled N-linked glycans is described. The HILIC column packed with a 1.7 mu m amide sorbent improves the peak capacity compared to a 3.0 mu m HILIC Column by a similar degree as observed in reversed-phase Ultra-performance liquid chromatography (RP-UPLC). The results indicated that the optimal peak capacity was achieved at flow rate 0.2-0.5 mL/min. HILIC method transfer guidelines were shown to further enhance the resolution of glycans by changing initial gradient conditions, flow rate, Column temperature, and different column lengths. Additionally, excellent resolution can be achieved in the separation of 2-AB labeled glycans released from fetuin, RNase B, and human IgG with a rapid analysis time. (C) 2009 Elsevier B.V. All rights reserved.

引用

页码：403 / 408

页数：6

共 28 条

[1] HYDROPHILIC-INTERACTION CHROMATOGRAPHY FOR THE SEPARATION OF PEPTIDES, NUCLEIC-ACIDS AND OTHER POLAR COMPOUNDS [J].