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题名: Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS
作者: Duan, JC;  Liang, Z;  Yang, C;  Zhang, J;  Zhang, LH;  Zhang, WB;  Zhang, YK
通讯作者: 张玉奎
关键词: enzymatic reactor ;  LC-MS/MS ;  monolithic column ;  protein identification
刊名: PROTEOMICS
发表日期: 2006
DOI: 10.1002/pmic.200500234
卷: 6, 期:2, 页:412-419
收录类别: ISTP ;  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
类目[WOS]: Biochemical Research Methods ;  Biochemistry & Molecular Biology
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: A monolithic enzymatic microreactor was prepared in a fused-silica capillary by in situ polymerization of acrylamide, glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) in the presence of a binary porogenic mixture of dodecanol and cyclohexanol, followed by ammonia solution treatment, glutaraldehyde activation and trypsin modification. The choice of acrylamide as co-monomer was found useful to improve the efficiency of trypsin modification, thus, to increase the enzyme activity. The optimized microreactor offered very low back pressure, enabling the fast digestion of proteins flowing through the reactor. The performance of the monolithic microreactor was demonstrated with the digestion of cytochrome c at high flow rate. The digests were then characterized by CE and HPLC-MS/MS with the sequence coverage of 57.7%. The digestion efficiency was found over 230 times as high as that of the conventional method. in addition, for the first time, protein digestion carried out in a mixture of water and ACN was compared with the conventional aqueous reaction using MS/MS detection, and the former solution was found more compatible and more efficient for protein digestion.
关键词[WOS]: MASS-SPECTROMETRY ;  CAPILLARY-ELECTROPHORESIS ;  AFFINITY-CHROMATOGRAPHY ;  PROTEOLYTIC DIGESTION ;  TRYPSIN MICROREACTOR ;  PEPTIDE SEPARATION ;  ION-EXCHANGE ;  SOL-GEL ;  PERFORMANCE ;  CHIP
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000235207700003
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/93527
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, Dalian 116023, Peoples R China

Recommended Citation:
Duan, JC,Liang, Z,Yang, C,et al. Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS[J]. PROTEOMICS,2006,6(2):412-419.
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