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题名: 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
作者: Tao, Dingyin1, 2;  Sun, Liangliang1, 2;  Zhu, Guijie1;  Liang, Yu1, 2;  Liang, Zhen1;  Zhang, Lihua1;  Zhang, Yukui1
关键词: Protein pre-fractionation ;  Serially coupled microcolumn ;  Shotgun proteome analysis ;  mu RPLC-MS/MS
刊名: JOURNAL OF SEPARATION SCIENCE
发表日期: 2011
DOI: 10.1002/jssc.201000632
卷: 34, 期:1, 页:83-89
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: To 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.
关键词[WOS]: TANDEM MASS-SPECTROMETRY ;  FREE-FLOW ELECTROPHORESIS ;  LIQUID-PHASE SEPARATION ;  PREFRACTIONATION TECHNIQUES ;  MULTICOMPARTMENT ELECTROLYZER ;  IMMOBILINE MEMBRANES ;  GEL-ELECTROPHORESIS ;  SHOTGUN PROTEOMICS ;  YEAST PROTEOME ;  HUMAN PLASMA
语种: 英语
WOS记录号: WOS:000286452700013
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/142693
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.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:
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.
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