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题名: Integration of capillary isoelectric focusing with monolithic immobilized pH gradient, immobilized trypsin microreactor and capillary zone electrophoresis for on-line protein analysis
作者: Wang, Tingting1, 2;  Ma, Junfeng1;  Zhu, Guijie1;  Shan, Yichu1;  Liang, Zhen1;  Zhang, Lihua1;  Zhang, Yukui1
关键词: Capillary zone electrophoresis ;  Immobilized enzyme microreactor ;  Integrated platform ;  Monolithic immobilized pH gradient ;  Trypsin
刊名: JOURNAL OF SEPARATION SCIENCE
发表日期: 2010-10-01
DOI: 10.1002/jssc.201000324
卷: 33, 期:20, 页:3194-3200
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: An integrated platform consisting of protein separation by CIEF with monolithic immobilized pH gradient (M IPG) on line digestion by trypsin based immobilized enzyme microreactor (trypsin IMER) and peptide separation by CZE was established In such a platform a tee unit was used not only to connect M IPG CIEF column and trypsin IMER but also to supply adjustment buffer to improve the compatibility of protein separation and digestion Another interface was made by a Teflon tube with a nick to couple IMER and CZE via a short capillary which was immerged in a centrifuge tube filled with 20 mmol/L glutamic acid to exchange protein digests buffer and keep electric contact for peptide separation By such a platform under the optimal conditions a mixture of ribonuclease A myoglobin and BSA was separated into 12 fractions by M IPG CIEF followed by on line digestion by trypsin IMER and peptide separation by CZE Many peaks of tryptic peptides corresponding to different proteins were observed with high UV signals indicating the excellent performance of such an integrated system We hope that the CE based on line platform developed herein would provide another powerful alternative for an integrated analysis of proteins
关键词[WOS]: TOP-DOWN ;  MASS-SPECTROMETRY ;  PROTEOMIC ANALYSIS ;  ENZYME REACTORS ;  SEPARATION ;  DIGESTION ;  CHROMATOGRAPHY ;  BOTTOM ;  MS ;  CE
语种: 英语
WOS记录号: WOS:000284181900012
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/142243
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 Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing, Peoples R China

Recommended Citation:
Wang, Tingting,Ma, Junfeng,Zhu, Guijie,et al. Integration of capillary isoelectric focusing with monolithic immobilized pH gradient, immobilized trypsin microreactor and capillary zone electrophoresis for on-line protein analysis[J]. JOURNAL OF SEPARATION SCIENCE,2010,33(20):3194-3200.
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