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学科主题: 物理化学
题名: Integrated Sample Pretreatment System for N-Linked Glycosylation Site Profiling with Combination of Hydrophilic Interaction Chromatography and PNGase F Immobilized Enzymatic Reactor via a Strong Cation Exchange Precolumn
作者: Qu, Yanyan1, 2;  Xia, Simin1, 2;  Yuan, Huiming1;  Wu, Qi1, 2;  Li, Man3;  Zou, Lijuan4;  Zhang, Lihua1;  Liang, Zhen1;  Zhang, Yukui1
通讯作者: 张丽华
刊名: ANALYTICAL CHEMISTRY
发表日期: 2011-10-01
DOI: 10.1021/ac201665e
卷: 83, 期:19, 页:7457-7463
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 1,1
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
摘要: Integrated Sample Pretreatment System for N-Linked Glycosylation Site Profiling with Combination of Hydrophilic Interaction Chromatography and PNGase F Immobilized Enzymatic Reactor via a Strong Cation Exchange Precolumn
研究领域[WOS]: Chemistry
英文摘要: An integrated sample pretreatment system, composed of a click maltose hydrophilic interaction chromatography (HILIC) column, a strong cation exchange (SCX) precolumn, and a PNGase F immobilized enzymatic reactor (IMER), was established for the simultaneous glycopeptide enrichment, sample buffer exchange, and online deglycosylation, by which the sample pretreatment for glycoproteome could be performed online automatically, beneficial to improve the efficiency and sensitivity of the N-linked glycosylation site identification. With such a system, the deglycosylated glycopeptide from the digests of avidin with the coexistence of 50 times (mass ratio) BSA could be selectively detected, and the detection limit as low as 5 fmol was achieved. Moreover, the sample pretreatment time was significantly shortened to similar to 1 h. Such a system was further successfully applied for analyzing the digest of the soluble fraction extracted from rat brain. A total of 120 unique glycoprotein groups and 196 N-linked glycosylation sites were identified by nanoreversed phase liquid chromatography-electrospray ionization-tandem mass spectrometry (nanoRPLC-ESI-MS/MS), with the injected digests amount as 6 mu g. All these results demonstrate that the integrated system is of great promise for N-linked glycosylation site profiling and could be further online coupled with nanoHPLC-ESI-MS/MS to achieve high-throughput glycoproteome analysis.
关键词[WOS]: INTERACTION LIQUID-CHROMATOGRAPHY ;  MASS-SPECTROMETRY ;  GLYCOPROTEINS ;  GLYCOPEPTIDES ;  IDENTIFICATION ;  ENRICHMENT ;  GLYCANS ;  DEGLYCOSYLATION ;  SEPARATION ;  PROTEINS
语种: 英语
WOS记录号: WOS:000295303600033
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115302
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
3.Dalian Med Univ, Affiliated Hosp 2, Dept Med Oncol, Dalian 116023, Peoples R China
4.Dalian Med Univ, Affiliated Hosp 2, Dept Radiat Oncol, Dalian 116023, Peoples R China

Recommended Citation:
Qu, Yanyan,Xia, Simin,Yuan, Huiming,et al. Integrated Sample Pretreatment System for N-Linked Glycosylation Site Profiling with Combination of Hydrophilic Interaction Chromatography and PNGase F Immobilized Enzymatic Reactor via a Strong Cation Exchange Precolumn[J]. ANALYTICAL CHEMISTRY,2011,83(19):7457-7463.
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