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题名: Vinyl functionalized silica hybrid monolith-based trypsin microreactor for on line digestion and separation via thiol-ene "click" strategy
作者: Chen, Yingzhuang1, 3;  Wu, Minghuo2;  Wang, Keyi1;  Chen, Bo3;  Yao, Shouzhuo1, 3;  Zou, Hanfa2;  Nie, Lihua1
关键词: Vinyl functionalized ;  Silica hybrid monolith ;  Enzyme microreactor ;  On-line digestion and separation ;  Thiol-ene "click" reaction
刊名: JOURNAL OF CHROMATOGRAPHY A
发表日期: 2011-11-04
DOI: 10.1016/j.chroma.2011.09.002
卷: 1218, 期:44, 页:7982-7988
收录类别: SCI
文章类型: Article
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
英文摘要: A novel thiol-ene "click" strategy for the preparation of monolithic trypsin microreactor was proposed. The hybrid organic-inorganic monolithic capillary column with ene-functionality was fabricated by sol-gel process using tetramethoxysilane (TMOS) and gamma-methacryloxypropyltrimethoxysilane (gamma-MAPS) as precursors. The disulfide bonds of trypsin were reduced to form free thiol groups. Then the trypsin containing free thiol groups was attached on the gamma-MAPS hybrid monolithic column with ene-functionality via thiol-ene click chemistry to form a trypsin microreactor. The activity of the trypsin microreactor was characterized by detecting the substrate (N alpha-p-tosyl-L-arginine methyl ester hydrochloride. TAME) and the product (N alpha-p-tosyl-L-arginine, TA) with on-line capillary zone electrophoresis. After investigating various synthesizing conditions, it was found that the microreactor with poly(N,N'-methylenebisacrylamide) as spacer can deliver the highest activity, yielding a rapid reaction rate. After repeatedly sampling and analyzing for 100 times, the monolithic trypsin microreactor still remained 87.5% of its initial activity. It was demonstrated that thiol-ene "click" strategy for the construction of enzyme microreactor is a promising method for the highly selective immobilization of proteins under mild conditions, especially enzymes with free thiol radicals. (c) 2011 Elsevier B.V. All rights reserved.
关键词[WOS]: INDUCED FLUORESCENCE DETECTION ;  IMMOBILIZED ENZYME REACTORS ;  CAPILLARY-ELECTROPHORESIS ;  PROTEOME ANALYSIS ;  CHEMISTRY ;  HYDROGELS ;  MS/MS ;  SPECTROMETRY ;  REACTIVITY ;  SURFACE
语种: 英语
WOS记录号: WOS:000296549000010
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/142290
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.Hunan Univ, Coll Chem & Chem Engn, State Key Lab Chemo Biosensing & Chemometr, Changsha 410082, Hunan, Peoples R China
2.CAS, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
3.Hunan Normal Univ, Minist Educ, Key Lab Chem Biol & Tradit Chinese Med Res, Changsha 410081, Hunan, Peoples R China

Recommended Citation:
Chen, Yingzhuang,Wu, Minghuo,Wang, Keyi,et al. Vinyl functionalized silica hybrid monolith-based trypsin microreactor for on line digestion and separation via thiol-ene "click" strategy[J]. JOURNAL OF CHROMATOGRAPHY A,2011,1218(44):7982-7988.
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