DICP OpenIR
Glycoproteomic Reactor for Human Plasma
Zhou, Hu; Hou, Weimin2; Denis, Nicholas J.; Zhou, Houjiang3; Vasilescu, Julian; Zou, Hanfa3; Figeys, Daniel1,2; Zou HF(邹汉法); Daniel Figeys
KeywordGlycoproteins Mass Spectrometry Proteomics Plasma Proteomic Reactor Microscale
Source PublicationJOURNAL OF PROTEOME RESEARCH
2009-02-01
ISSN1535-3893
DOI10.1021/pr800734r
Volume8Issue:2Pages:556-566
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank3;4
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordN-LINKED GLYCOPROTEINS ; MULTILECTIN AFFINITY-CHROMATOGRAPHY ; MASS-SPECTROMETRY ; PROTEOMIC REACTOR ; HYDRAZIDE CHEMISTRY ; GLYCOSYLATION SITES ; SERUM-PROTEINS ; IDENTIFICATION ; CAPTURE ; CANCER
AbstractWe 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.
Language英语
URL查看原文
WOS IDWOS:000263193300016
Citation statistics
Cited Times:27[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/101515
Collection中国科学院大连化学物理研究所
Corresponding AuthorZou HF(邹汉法); Daniel Figeys
Affiliation1.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
GB/T 7714
Zhou, Hu,Hou, Weimin,Denis, Nicholas J.,et al. Glycoproteomic Reactor for Human Plasma[J]. JOURNAL OF PROTEOME RESEARCH,2009,8(2):556-566.
APA Zhou, Hu.,Hou, Weimin.,Denis, Nicholas J..,Zhou, Houjiang.,Vasilescu, Julian.,...&Daniel Figeys.(2009).Glycoproteomic Reactor for Human Plasma.JOURNAL OF PROTEOME RESEARCH,8(2),556-566.
MLA Zhou, Hu,et al."Glycoproteomic Reactor for Human Plasma".JOURNAL OF PROTEOME RESEARCH 8.2(2009):556-566.
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