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题名: Glycoproteomic Reactor for Human Plasma
作者: Zhou, Hu;  Hou, Weimin2;  Denis, Nicholas J.;  Zhou, Houjiang3;  Vasilescu, Julian;  Zou, Hanfa3;  Figeys, Daniel1, 2
通讯作者: 邹汉法 ;  Daniel Figeys
关键词: glycoproteins ;  mass spectrometry ;  proteomics ;  plasma ;  proteomic reactor ;  microscale
刊名: JOURNAL OF PROTEOME RESEARCH
发表日期: 2009-02-01
DOI: 10.1021/pr800734r
卷: 8, 期:2, 页:556-566
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1809
产权排名: 3;4
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: We describe the development of a glycoproteomic reactor that combines multiple biochemical and chemical protein processing into a single device for the study of N-glycosylated proteins. The glycoproteins are first enriched by concanavalin A affinity chromatography and then transferred onto and efficiently processed in the glycoproteomic reactor. This glycoproteomic reactor combines protein concentration and purification, disulfide bond reduction, peptide-N-glycosidase-mediated (18)O-labeling and deglycosylation, alkylation, tryptic digestion and pH based fractionation in a device that has an interstitial volume (reaction volume) of similar to 1 mu L. We demonstrated the potential of the glycoproteomic reactor using human plasma. Under stringent criteria, 82 unique glycopeptides representing 41 unique glycoproteins were identified from as little as 5 mu L of human plasma. Our glycoproteomic reactor reduces the sample processing time to less than 1.5 h, reduces the reagent consumption while providing over 1000-fold concentration of the sample, provides efficient removal of high concentration of glycan buffer, and, finally, allows both glycopeptides and nonglycosylated tryptic peptides to be analyzed by the mass spectrometer which provides much greater protein coverage and more reliable identifications.
关键词[WOS]: N-LINKED GLYCOPROTEINS ;  MULTILECTIN AFFINITY-CHROMATOGRAPHY ;  MASS-SPECTROMETRY ;  PROTEOMIC REACTOR ;  HYDRAZIDE CHEMISTRY ;  GLYCOSYLATION SITES ;  SERUM-PROTEINS ;  IDENTIFICATION ;  CAPTURE ;  CANCER
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000263193300016
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/101515
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.Univ Ottawa, Ottawa Inst Syst Biol, Dept Biochem Microbiol & Immunol, Fac Med, Ottawa, ON K1H 8M5, Canada
2.Univ Ottawa, Fac Sci, Dept Chem, Ottawa, ON K1N 6N5, Canada
3.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Dalian, Peoples R China

Recommended Citation:
Zhou, Hu,Hou, Weimin,Denis, Nicholas J.,et al. Glycoproteomic Reactor for Human Plasma[J]. JOURNAL OF PROTEOME RESEARCH,2009,8(2):556-566.
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