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Vinyl functionalized silica hybrid monolith-based trypsin microreactor for on line digestion and separation via thiol-ene "click" strategy
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
2011-11-04
DOI10.1016/j.chroma.2011.09.002
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
关键词[WOS]INDUCED FLUORESCENCE DETECTION ; IMMOBILIZED ENZYME REACTORS ; CAPILLARY-ELECTROPHORESIS ; PROTEOME ANALYSIS ; CHEMISTRY ; HYDROGELS ; MS/MS ; SPECTROMETRY ; REACTIVITY ; SURFACE
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.
语种英语
WOS记录号WOS:000296549000010
引用统计
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/142290
专题中国科学院大连化学物理研究所
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
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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.
APA Chen, Yingzhuang.,Wu, Minghuo.,Wang, Keyi.,Chen, Bo.,Yao, Shouzhuo.,...&Nie, Lihua.(2011).Vinyl functionalized silica hybrid monolith-based trypsin microreactor for on line digestion and separation via thiol-ene "click" strategy.JOURNAL OF CHROMATOGRAPHY A,1218(44),7982-7988.
MLA Chen, Yingzhuang,et al."Vinyl functionalized silica hybrid monolith-based trypsin microreactor for on line digestion and separation via thiol-ene "click" strategy".JOURNAL OF CHROMATOGRAPHY A 1218.44(2011):7982-7988.
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