DICP OpenIR
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
KeywordIon Exchange Membrane Enzyme Micro-reactor Protein Analysis Liquid Chromatography Mass Spectrometry Lc-ms
Source PublicationANALYTICAL AND BIOANALYTICAL CHEMISTRY
2012-04-01
ISSN1618-2642
DOI10.1007/s00216-012-5812-2
Volume403Issue:1Pages:239-246
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordIMMOBILIZED TRYPSIN MICROREACTOR ; PEPTIDE SEPARATION ; ESI-MS ; DIGESTION ; IDENTIFICATION ; CAPILLARY ; COLUMN ; MS/MS
AbstractIn 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. '
Language英语
WOS IDWOS:000301839700018
PublisherSPRINGER HEIDELBERG
Citation statistics
Cited Times:9[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/167287
Collection中国科学院大连化学物理研究所
Corresponding AuthorLiu, Huwei
AffiliationPeking 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
Recommended Citation
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]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2012,403(1):239-246.
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.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,403(1),239-246.
MLA Zhou, Zhigui,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.
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