中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis

文献类型:期刊论文

作者Zhang, Jialing; Zhou, Zhigui; Yang, Youyou; Zhang, Zhengxiang; Bai, Yu; Liao, Yiping; Liu, Huwei
刊名ANALYTICAL AND BIOANALYTICAL CHEMISTRY
出版日期2012-04-01
卷号403期号:1页码:239-246
关键词Ion Exchange Membrane Enzyme Micro-reactor Protein Analysis Liquid Chromatography Mass Spectrometry Lc-ms
ISSN号1618-2642
DOI10.1007/s00216-012-5812-2
文献子类Article
英文摘要In this article, we developed a membrane-based enzyme micro-reactor by directly using commercial polystyrene-divinylbenzene cation-exchange membrane as the support for trypsin immobilization via electrostatic and hydrophobic interactions and successfully applied it for protein digestion. The construction of the reactor can be simply achieved by continuously pumping trypsin solution through the reactor for only 2 min, which was much faster than the other enzyme immobilization methods. In addition, the membrane reactor could be rapidly regenerated within 35 min, resulting in a "new" reactor for the digestion of every protein sample, completely eliminating the cross-interference of different protein samples. The amount and the activity of immobilized trypsin were measured, and the repeatability of the reactor was tested, with an RSD of 3.2% for the sequence coverage of cytochrome c in ten digestion replicates. An integrated platform for protein analysis, including online protein digestion and peptide separation and detection, was established by coupling the membrane enzyme reactor with liquid chromatography-quadrupole time-of-flight mass spectrometry. The performance of the platform was evaluated using cytochrome c, myoglobin, and bovine serum albumin, showing that even in the short digestion time of several seconds the obtained sequence coverages was comparable to or higher than that with in-solution digestion. The system was also successfully used for the analysis of proteins from yeast cell lysate. '
WOS关键词IMMOBILIZED TRYPSIN MICROREACTOR ; PEPTIDE SEPARATION ; ESI-MS ; DIGESTION ; IDENTIFICATION ; CAPILLARY ; COLUMN ; MS/MS
WOS研究方向Biochemistry & Molecular Biology ; Chemistry
语种英语
WOS记录号WOS:000301839700018
出版者SPRINGER HEIDELBERG
源URL[http://cas-ir.dicp.ac.cn/handle/321008/167286]  
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
通讯作者Liu, Huwei
作者单位Peking Univ, Beijing Natl Lab Mol Sci, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Inst Analyt Chem,Coll Chem & Mol Engn, Beijing 100871, Peoples R China
推荐引用方式
GB/T 7714
Zhang, Jialing,Zhou, Zhigui,Yang, Youyou,et al. Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2012,403(1):239-246.
APA Zhang, Jialing.,Zhou, Zhigui.,Yang, Youyou.,Zhang, Zhengxiang.,Bai, Yu.,...&Liu, Huwei.(2012).Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,403(1),239-246.
MLA Zhang, Jialing,et al."Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 403.1(2012):239-246.

入库方式: OAI收割

来源:大连化学物理研究所

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