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学科主题: 物理化学
题名: Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis
作者: Liang, Yu1, 2;  Tao, Dingyin1, 2;  Ma, Junfeng1;  Sun, Liangliang1, 2;  Liang, Zhen1;  Zhang, Lihua1;  Zhang, Yukui1
通讯作者: 张丽华
关键词: Immobilized enzyme reactor ;  Hydrophilic monolith ;  Capillary ;  Microchip ;  Proteomic analysis
刊名: JOURNAL OF CHROMATOGRAPHY A
发表日期: 2011-05-20
DOI: 10.1016/j.chroma.2011.02.073
卷: 1218, 期:20, 页:2898-2905
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 ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
摘要: Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: A novel kind of hydrophilic monolith based immobilized enzyme reactors (IMERs) was prepared both in UV-transparent capillaries and on glass microchips by the photopolymerization of N-acryloxysuccinimide and poly(ethylene glycol)diacrylate, followed by trypsin immobilization. The performance of capillary IMERs for protein digestion was evaluated by the digestion of myoglobin with the residential time from 12s to 71 s. With mu RPLC-ESI-MS/MS analysis, the obtained sequence coverages were all over 80%, comparable to that obtained by in-solution digestion for 12 h. The nonspecific absorption of BSA on monolithic support was evaluated, and no obvious protein residue was observed by a fluorescence assay. Moreover, no carry-over of the digests on the capillary IMER was found after the digestion of myoglobin (24 mu g) and BSA (9 mu g), which further demonstrated the good hydrophilicity of such matrix. In addition, an integrated microchip-based system involving on-line protein digestion by microchip-based IMER, peptides separation by nanoRPLC and identification by ESI-MS/MS was established, by which a mixture of standard proteins and one RPLC fraction of Escherichia coli extract were successfully identified, indicating that the hydrophilic monolith based IMER might provide a promising tool for high-throughput proteomic analysis. (C) 2011 Elsevier B.V. All rights reserved.
关键词[WOS]: POROUS POLYMER MONOLITHS ;  MASS-SPECTROMETRY ;  PROTEIN IDENTIFICATION ;  ELECTROSPRAY-IONIZATION ;  LIQUID-CHROMATOGRAPHY ;  SOL-GEL ;  TRYPSIN ;  SEPARATION ;  DIGESTION ;  CHIP
语种: 英语
WOS记录号: WOS:000291065500010
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115332
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, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China

Recommended Citation:
Liang, Yu,Tao, Dingyin,Ma, Junfeng,et al. Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis[J]. JOURNAL OF CHROMATOGRAPHY A,2011,1218(20):2898-2905.
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