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Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis
Tao, Dingyin1,2; Zhu, Guajie1,2; Sun, Liangliang1,2; Ma, Junfeng1,2; Liang, Zhen1; Zhang, Weibing1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
KeywordMu Rplc Ms/ms Serially Coupled Microcolumn Shotgun Proteome Analysis
Source PublicationPROTEOMICS
2009-04-01
DOI10.1002/pmic.200800649
Volume9Issue:7Pages:2029-2036
Indexed BySCI
SubtypeArticle
Department18
Funding Project1810
Contribution Rank1;1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology
WOS Research AreaBiochemistry & Molecular Biology
WOS Keyword2-DIMENSIONAL GEL-ELECTROPHORESIS ; MONOLITHIC SILICA COLUMNS ; TANDEM MASS-SPECTROMETRY ; LOW-ABUNDANCE PROTEINS ; ESCHERICHIA-COLI ; PEAK-CAPACITY ; PERFORMANCE ; TEMPERATURE ; TECHNOLOGY ; AUTOMATION
AbstractMicrocolumn RPLC (mu RPLC) is one of the optimum separation modes for shotgun proteomic analysis. To identify as many proteins as possible by MS/MS, the improvement on separation efficiency and peak capacity of mu RPLC is indispensable. Although the increase in column length is one of the effective solutions, the preparation of along microcolumn is rather difficult due to the high backpressure generated during the packing procedure. In our recent work, through connecting microcolumns of 5, 10, and 15 cm length via unions with minimal dead volume, long microcolumns with length up to 30 cm were obtained, with which 318 proteins were identified from proteins extracted from Escherichia coli by mu RPLC-ESI MS/MS, and similar distributions of M(w) and pI were found with single and various coupled microcolumns. Furthermore, by using MS/MS with improved sensitivity, with such a serially coupled 30 cm long microcolumn, 1692 proteins were identified within 7 h from rat brain tissue, with false positive rate (FPR) < 1%. All these results demonstrated that serially couple microcolumns might be of great promising to improve the separation capacity of mu RPLC in shotgun proteomic analysis.
Language英语
URL查看原文
WOS IDWOS:000265339000025
Citation statistics
Cited Times:14[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/101655
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang LH(张丽华)
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
Recommended Citation
GB/T 7714
Tao, Dingyin,Zhu, Guajie,Sun, Liangliang,et al. Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis[J]. PROTEOMICS,2009,9(7):2029-2036.
APA Tao, Dingyin.,Zhu, Guajie.,Sun, Liangliang.,Ma, Junfeng.,Liang, Zhen.,...&张丽华.(2009).Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis.PROTEOMICS,9(7),2029-2036.
MLA Tao, Dingyin,et al."Serially coupled microcolumn reversed phase liquid chromatography for shotgun proteomic analysis".PROTEOMICS 9.7(2009):2029-2036.
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