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学科主题: 物理化学
题名: Integrated SDS removal and protein digestion by hollow fiber membrane based device for SDS-assisted proteome analysis
作者: Xia, Simin1, 2;  Yuan, Huiming1;  Chen, Yuanbo1, 2;  Liang, Zheng1;  Zhang, Lihua1;  Zhang, Yukui1
通讯作者: 张丽华
关键词: SDS removal ;  On-line digestion ;  Hollow fiber membrane interface ;  Immobilized enzymatic reactor ;  Proteome analysis
刊名: TALANTA
发表日期: 2015-08-15
DOI: 10.1016/j.talanta.2015.04.011
卷: 141, 期:1, 页:235-238
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: In this work, a novel integrated sample preparation device for SDS-assisted proteome analysis was developed, by which proteins dissolved in 4% (w/v) SDS were first diluted by 50% methanol, and then SDS was online removed by a hollow fiber membrane interface (HFMI) with 50 mM ammonium bicarbonate (pH 8.0) as an exchange buffer, finally digested by an immobilized enzyme reactor (IMER). To evaluate the performance of such an integrated device, bovine serum albumin dissolved in 4% (w/v) SDS as a model sample was analyzed; it could be found that similar to that obtained by direct analysis of BSA digests without SDS (the sequence coverage of 60.3 +/- 1.0%, n=3), with HFMI as an interface for SDS removal, BSA was identified with the sequence coverage of 61.0 +/- 1.0% (n=3). However, without SDS removal by HFMI, BSA could not be digested by the IMER and none peptides could be detected. In addition, such an integrated sample preparation device was also applied for the analysis of SDS extracted proteins from rat brain, compared to those obtained by filter-aided sample preparation (FASP), not only the identified protein group and unique peptide number were increased by 12% and 39% respectively, but also the sample pretreatment time was shortened from 24 h to 4 h. All these results demonstrated that such an integrated sample preparation device would provide an alternative tool for SDS assisted proteome analysis. (C) 2015 Elsevier B.V. All rights reserved.
关键词[WOS]: SAMPLE PREPARATION METHOD ;  TANDEM MASS-SPECTROMETRY ;  LIQUID-CHROMATOGRAPHY ;  GEL-ELECTROPHORESIS ;  PEPTIDE SEPARATION ;  IDENTIFICATION
语种: 英语
WOS记录号: WOS:000356987500037
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/143709
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, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100039, Peoples R China

Recommended Citation:
Xia, Simin,Yuan, Huiming,Chen, Yuanbo,et al. Integrated SDS removal and protein digestion by hollow fiber membrane based device for SDS-assisted proteome analysis[J]. TALANTA,2015,141(1):235-238.
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