Ion-exchange-membrane-based enzyme micro-reactor coupled online with   liquid chromatography-mass spectrometry for protein analysis

doi:10.1007/s00216-012-5812-2

CORC > 北京大学 > 化学与分子工程学院

	Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis
	Zhou, Zhigui ; Yang, Youyou ; Zhang, Jialing ; Zhang, Zhengxiang ; Bai, Yu ; Liao, Yiping ; Liu, Huwei
刊名	分析化学和生物分析化学
	2012
关键词	Ion exchange membrane Enzyme micro-reactor Protein analysis Liquid chromatography Mass spectrometry LC-MS IMMOBILIZED TRYPSIN MICROREACTOR PEPTIDE SEPARATION ESI-MS DIGESTION IDENTIFICATION CAPILLARY COLUMN MS/MS
DOI	10.1007/s00216-012-5812-2
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. '; http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000301839700018&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=8e1609b174ce4e31116a60747a720701 ; Biochemical Research Methods; Chemistry, Analytical; SCI(E); EI; PubMed; 5; ARTICLE; 1; 239-246; 403
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/149996]
专题	化学与分子工程学院
推荐引用方式 GB/T 7714	Zhou, Zhigui,Yang, Youyou,Zhang, Jialing,et al. Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis[J]. 分析化学和生物分析化学,2012.
APA	Zhou, Zhigui.,Yang, Youyou.,Zhang, Jialing.,Zhang, Zhengxiang.,Bai, Yu.,...&Liu, Huwei.(2012).Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis.分析化学和生物分析化学.
MLA	Zhou, Zhigui,et al."Ion-exchange-membrane-based enzyme micro-reactor coupled online with liquid chromatography-mass spectrometry for protein analysis".分析化学和生物分析化学 (2012).