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Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS
Duan, JC; Liang, Z; Yang, C; Zhang, J; Zhang, LH; Zhang, WB; Zhang, YK; Zhang YK(张玉奎); Zhang YK(张玉奎)
关键词Enzymatic Reactor Lc-ms/ms Monolithic Column Protein Identification
刊名PROTEOMICS
2006
DOI10.1002/pmic.200500234
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
关键词[WOS]MASS-SPECTROMETRY ; CAPILLARY-ELECTROPHORESIS ; AFFINITY-CHROMATOGRAPHY ; PROTEOLYTIC DIGESTION ; TRYPSIN MICROREACTOR ; PEPTIDE SEPARATION ; ION-EXCHANGE ; SOL-GEL ; PERFORMANCE ; CHIP
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.
语种英语
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WOS记录号WOS:000235207700003
引用统计
被引频次:75[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/93527
专题中国科学院大连化学物理研究所
通讯作者Zhang YK(张玉奎); Zhang YK(张玉奎)
作者单位Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China
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GB/T 7714
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.
APA Duan, JC.,Liang, Z.,Yang, C.,Zhang, J.,Zhang, LH.,...&张玉奎.(2006).Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS.PROTEOMICS,6(2),412-419.
MLA Duan, JC,et al."Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS".PROTEOMICS 6.2(2006):412-419.
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