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学科主题物理化学
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
Qu, Yanyan1,2; Xia, Simin1,2; Yuan, Huiming1; Wu, Qi1,2; Li, Man3; Zou, Lijuan4; Zhang, Lihua1; Liang, Zhen1; Zhang, Yukui1; Zhang LH(张丽华)
刊名ANALYTICAL CHEMISTRY
2011-10-01
ISSN待补充
DOI10.1021/ac201665e
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
研究领域[WOS]Chemistry
关键词[WOS]INTERACTION LIQUID-CHROMATOGRAPHY ; MASS-SPECTROMETRY ; GLYCOPROTEINS ; GLYCOPEPTIDES ; IDENTIFICATION ; ENRICHMENT ; GLYCANS ; DEGLYCOSYLATION ; SEPARATION ; PROTEINS
英文摘要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; 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.
语种英语
WOS记录号WOS:000295303600033
引用统计
被引频次:46[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/115302
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
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
推荐引用方式
GB/T 7714
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.
APA Qu, Yanyan.,Xia, Simin.,Yuan, Huiming.,Wu, Qi.,Li, Man.,...&张丽华.(2011).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.ANALYTICAL CHEMISTRY,83(19),7457-7463.
MLA Qu, Yanyan,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".ANALYTICAL CHEMISTRY 83.19(2011):7457-7463.
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