Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis

	Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis
	Liang, Yu 1,2; Tao, Dingyin 1,2; Ma, Junfeng 1; Sun, Liangliang 1,2; Liang, Zhen 1; Zhang, Lihua 1; Zhang, Yukui 1
刊名	journal of chromatography a
	2011-05-20
卷号	1218 期号:20 页码:2898-2905
关键词	Immobilized enzyme reactor Hydrophilic monolith Capillary Microchip Proteomic analysis
ISSN号	待补充
通讯作者	张丽华
产权排序	1,1
中文摘要	hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis
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.
学科主题	物理化学
WOS标题词	science & technology ; life sciences & biomedicine ; physical sciences
类目[WOS]	biochemical research methods ; chemistry, analytical
研究领域[WOS]	biochemistry & molecular biology ; chemistry
关键词[WOS]	porous polymer monoliths ; mass-spectrometry ; protein identification ; electrospray-ionization ; liquid-chromatography ; sol-gel ; trypsin ; separation ; digestion ; chip
收录类别	SCI
语种	英语
WOS记录号	WOS:000291065500010
公开日期	2012-07-09
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/115332]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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
推荐引用方式 GB/T 7714	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.
APA	Liang, Yu.,Tao, Dingyin.,Ma, Junfeng.,Sun, Liangliang.,Liang, Zhen.,...&Zhang, Yukui.(2011).Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis.journal of chromatography a,1218(20),2898-2905.
MLA	Liang, Yu,et al."Hydrophilic monolith based immobilized enzyme reactors in capillary and on microchip for high-throughput proteomic analysis".journal of chromatography a 1218.20(2011):2898-2905.