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题名: Integrated Platform for Proteome Analysis with Combination of Protein and Peptide Separation via Online Digestion
作者: Yuan, Huiming1, 2;  Zhang, Lihua1;  Hou, Chunyan1, 2;  Zhu, Guijie1, 2;  Tao, Dingyin1, 2;  Liang, Zhen1;  Zhang, Yukui1
通讯作者: 张丽华
刊名: ANALYTICAL CHEMISTRY
发表日期: 2009-11-01
DOI: 10.1021/ac900310y
卷: 81, 期:21, 页:8708-8714
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 1;1
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: An integrated platform with the combination of protein and peptide separation was established via online protein digestion, by which proteins were first separated by a microcolumn packed with mixed weak anion and weak cation exchange (WAX/WCX) particles under a series of salt steps, online digested by a trypsin immobilized microenzymatic reactor (IMER), trapped and desalted by two parallel CS precolumns, separated by microreversed-phase liquid chromatography (mu RPLC) under a linear gradient of organic modifier concentration, and finally identified by electrospray ionization-MS/MS (ESI-MS/MS). To evaluate the performance of such a platform, a mixture of myoglobin, cytochrome c, bovine serum albumin (BSA), and alpha-casein, with mass ranging from 25 ng to 2 mu g, was analyzed. Compared to the methods by offline protein fractionation and shotgun based strategy, the analysis time, including sample preparation, digestion, desalting, separation, and detection, was shortened from ca. 30 to 5 h, and cytochrome c with abundance of 25 ng could be identified with improved sequence coverage. Furthermore, such an integrated platform was successfully applied into the analysis of proteins extracted from human lung cancer cells. Compared with the results obtained by the shotgun approach, the identified protein number was increased by 30%. All these results demonstrated that such an integrated approach would be an attractive alternative to commonly applied approaches for proteome research.
关键词[WOS]: MULTIDIMENSIONAL LIQUID-CHROMATOGRAPHY ;  TANDEM MASS-SPECTROMETRY ;  TOP-DOWN PROTEOMICS ;  ION-EXCHANGE ;  IDENTIFICATION ;  MICROREACTOR ;  BOTTOM ;  SYSTEM ;  COLUMN ;  MONOLITHS
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000276191900013
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/102393
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 100039, Peoples R China

Recommended Citation:
Yuan, Huiming,Zhang, Lihua,Hou, Chunyan,et al. Integrated Platform for Proteome Analysis with Combination of Protein and Peptide Separation via Online Digestion[J]. ANALYTICAL CHEMISTRY,2009,81(21):8708-8714.
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