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Highly efficient proteome analysis with combination of protein pre-fractionation by preparative microscale solution isoelectric focusing and identification by mu RPLC-MS/MS with serially coupled long microcolumn
Tao, Dingyin1,2; Sun, Liangliang1,2; Zhu, Guijie1; Liang, Yu1,2; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1
KeywordProtein Pre-fractionation Serially Coupled Microcolumn Shotgun Proteome Analysis Mu Rplc-ms/ms
Source PublicationJOURNAL OF SEPARATION SCIENCE
2011
DOI10.1002/jssc.201000632
Volume34Issue:1Pages:83-89
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordTANDEM MASS-SPECTROMETRY ; FREE-FLOW ELECTROPHORESIS ; LIQUID-PHASE SEPARATION ; PREFRACTIONATION TECHNIQUES ; MULTICOMPARTMENT ELECTROLYZER ; IMMOBILINE MEMBRANES ; GEL-ELECTROPHORESIS ; SHOTGUN PROTEOMICS ; YEAST PROTEOME ; HUMAN PLASMA
AbstractTo improve the efficiency of proteome analysis, a strategy with the combination of protein pre-fractionation by preparative microscale solution isoelectric focusing, peptide separation by mu RPLC with serially coupled long microcolumn and protein identification by ESI-MS/MS was proposed. By preparative microscale solution isoelectric focusing technique, proteins extracted from whole cell lysates of Escherichia coli were fractionated into five chambers divided by isoelectric membranes, respectively with pH range from 3.0 to 4.6, 4.6 to 5.4, 5.4 to 6.2, 6.2 to 7.0 and 7.0 to 10.0. Compared to the traditional on-gel IFF, the protein recovery could be obviously improved to over 95%. Subsequently, the enriched and fractionated proteins in each chamber were digested, and further separated by a 30-cm long serially coupled RP microcolumn. Through the detection by ESI-MS/MS, about 200 proteins were identified in each fraction, and in total 835 proteins were identified even with one-dimensional mu RPLC-MS/MS system. All these results demonstrate that by such a combination strategy, highly efficient proteome analysis could be achieved, not only due to the in-solution protein enrichment and pre-fractionation with improved protein recovery but also owing to the increased separation capacity of serially coupled long mu RPLC columns.
Language英语
WOS IDWOS:000286452700013
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Cited Times:6[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/142693
Collection中国科学院大连化学物理研究所
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,Sun, Liangliang,Zhu, Guijie,et al. Highly efficient proteome analysis with combination of protein pre-fractionation by preparative microscale solution isoelectric focusing and identification by mu RPLC-MS/MS with serially coupled long microcolumn[J]. JOURNAL OF SEPARATION SCIENCE,2011,34(1):83-89.
APA Tao, Dingyin.,Sun, Liangliang.,Zhu, Guijie.,Liang, Yu.,Liang, Zhen.,...&Zhang, Yukui.(2011).Highly efficient proteome analysis with combination of protein pre-fractionation by preparative microscale solution isoelectric focusing and identification by mu RPLC-MS/MS with serially coupled long microcolumn.JOURNAL OF SEPARATION SCIENCE,34(1),83-89.
MLA Tao, Dingyin,et al."Highly efficient proteome analysis with combination of protein pre-fractionation by preparative microscale solution isoelectric focusing and identification by mu RPLC-MS/MS with serially coupled long microcolumn".JOURNAL OF SEPARATION SCIENCE 34.1(2011):83-89.
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