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学科主题物理化学
Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis
Zhu, Jun; Wang, Fangjun; Chen, Rui; Cheng, Kai; Xu, Bo; Guo, Zhimou; Liang, Xinmiao; Ye, Mingliang; Zou, Hanfa; Zou HF(邹汉法); Ye ML(叶明亮)
刊名ANALYTICAL CHEMISTRY
2012-06-05
ISSN0003-2700
DOI10.1021/ac3000732
84期:11页:5146-5153
收录类别SCI
文章类型Article
部门归属18
项目归属1809
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]LECTIN AFFINITY-CHROMATOGRAPHY ; PHASE LIQUID-CHROMATOGRAPHY ; MASS-SPECTROMETRY ; PROTEOME ANALYSIS ; GLYCOSYLATION HETEROGENEITY ; ABERRANT GLYCOSYLATION ; MALDI-MS ; GLYCOPEPTIDES ; PLASMA ; IDENTIFICATION
英文摘要Sample handling procedures including protein digestion, glycopeptide enrichment, and deglycosylation have significant impact on the performance of glycoproteome analysis. Several glycoproteomic analysis systems were developed to integrate some of these sample preparation procedures. However, no microsystem integrates all of above three procedures together. In this work, we developed a glycoproteomic microreactor enabling seamless integration of all these procedures. In this reactor, trypsin digestion was accelerated by adding acetonitrile to 80%, and after acidification of protein digest by trifluoroacetic acid (TFA), the following hydrophilic interaction chromatography (HILIC) enrichment and deglycosylation were sequentially performed without any desalting, lyophilization, or buffer exchange steps. The total processing time could be as short as 1.5 h. The detection limit of human IgG as low as 30 fmol was also achieved. When applied to human serum glycoproteome analysis, a total number of 92, 178, and 221 unique N-glycosylation sites were identified from three replicate analyses of 10 nL, 100 nL, and 1 mu L of human serum, respectively. It was demonstrated that the glycoproteomic microreactor based method had very high sensitivity and was well suited for glycoproteome analysis of minute protein samples.
语种英语
WOS记录号WOS:000304783100073
引用统计
被引频次:73[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/321008/117706
专题中国科学院大连化学物理研究所
通讯作者Zou HF(邹汉法); Ye ML(叶明亮)
作者单位Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China
推荐引用方式
GB/T 7714
Zhu, Jun,Wang, Fangjun,Chen, Rui,et al. Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis[J]. ANALYTICAL CHEMISTRY,2012,84(11):5146-5153.
APA Zhu, Jun.,Wang, Fangjun.,Chen, Rui.,Cheng, Kai.,Xu, Bo.,...&叶明亮.(2012).Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis.ANALYTICAL CHEMISTRY,84(11),5146-5153.
MLA Zhu, Jun,et al."Centrifugation Assisted Microreactor Enables Facile Integration of Trypsin Digestion, Hydrophilic Interaction Chromatography Enrichment, and On-Column Deglycosylation for Rapid and Sensitive N-Glycoproteome Analysis".ANALYTICAL CHEMISTRY 84.11(2012):5146-5153.
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