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Monolithic Capillary Column Based Glycoproteomic Reactor for High-Sensitive Analysis of N-Glycoproteome
Liu, Jing1,2; Wang, Fangjun1; Lin, Hui1,2; Zhu, Jun1,2; Bian, Yangyang1,2; Cheng, Kai1,2; Zou, Hanfa1
Source PublicationANALYTICAL CHEMISTRY
2013-03-05
DOI10.1021/ac400315n
Volume85Issue:5Pages:2847-2852
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordALTERED GLYCOSYLATION ; MASS-SPECTROMETRY ; PROTEINS ; REVEALS ; CELLS
AbstractDespite the importance of protein N-glycosylation in a series of biological processes, in-depth characterization of protein glycosylation is still a challenge due to the high complexity of biological samples and the lacking of highly sensitive detection technologies. We developed a monolithic capillary column based glycoproteomic reactor enabling high. sensitive mapping of N-glycosylation sites from minute amounts of sample. Unlike the conventional proteomic reactors with only strong-cation exchange or hydrophilic-interaction chromatography columns, this novel glycoproteomic reactor was composed of an 8 cm long C12 hydrophobic monolithic capillary column for protein digestion and a 6 cm long organic-silica hybrid hydrophilic monolithic capillary column for glycopeptides enrichment and deglycosylation, which could complete whole-sample preparation including protein purification/desalting, tryptic digestion, enrichment, and deglycosylation of glycopeptides within about 3 h. The developed reactor exhibited high detection sensitivity in mapping of N-glycosylation sites by detection limit of horseradish peroxidase as low as 2.5 fmol. This reactor also demonstrated the ability in complex sample analysis, and in total, 486 unique N-glycosylation sites were reliably mapped in three replicate analyses of a protein sample extracted from similar to 10(4) HeLa cells.
Language英语
WOS IDWOS:000317031600043
Citation statistics
Cited Times:43[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/137787
Collection中国科学院大连化学物理研究所
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Liu, Jing,Wang, Fangjun,Lin, Hui,et al. Monolithic Capillary Column Based Glycoproteomic Reactor for High-Sensitive Analysis of N-Glycoproteome[J]. ANALYTICAL CHEMISTRY,2013,85(5):2847-2852.
APA Liu, Jing.,Wang, Fangjun.,Lin, Hui.,Zhu, Jun.,Bian, Yangyang.,...&Zou, Hanfa.(2013).Monolithic Capillary Column Based Glycoproteomic Reactor for High-Sensitive Analysis of N-Glycoproteome.ANALYTICAL CHEMISTRY,85(5),2847-2852.
MLA Liu, Jing,et al."Monolithic Capillary Column Based Glycoproteomic Reactor for High-Sensitive Analysis of N-Glycoproteome".ANALYTICAL CHEMISTRY 85.5(2013):2847-2852.
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