DICP OpenIR
Subject Area物理化学
Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith
Ma, Junfeng1,2; Hou, Chunyan1,2; Liang, Yu1,2; Wang, Tingting1,2; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
KeywordImmobilized Enzyme Reactor Metal-ion Chelation Organic-inorganic Hybrid Silica Monolith Protein Digestion Technology Trypsin
Source PublicationPROTEOMICS
2011-03-01
ISSN待补充
DOI10.1002/pmic.201000550
Volume11Issue:5Pages:991-995
Indexed BySCI
SubtypeArticle
Department18
Funding Project1810
Contribution Rank1,1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordMASS-SPECTROMETRY ; CAPILLARY MICROREACTOR ; PROTEIN IDENTIFICATION ; SOL-GEL ; TRYPSIN ; ONLINE ; ELECTROPHORESIS ; MICROSPHERES ; SEPARATION ; DIGESTION
AbstractEfficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith; A metal-ion chelate immobilized enzyme reactor (IMER) supported on organic-inorganic hybrid silica monolith was developed for rapid digestion of proteins. The monolithic support was in situ prepared in a fused silica capillary via the polycondensation between tetraethoxysilane hydrolytic sol and iminodiacetic acid conjugated glycidoxypropyltrimethoxysilane. After activated by Cu(2+), trypsin was immobilized onto the monolithic support via metal chelation. Proteolytic capability of such an IMER was evaluated by the digestion of myoglobin and BSA, and the digests were further analyzed by microflow reversed-phase liquid chromatography with ESI-MS/MS. Similar sequence coverages of myoglobin and BSA were obtained by IMER, in comparison to those obtained by in-solution digestion (91 versus 92% for 200 ng myoglobin, and 26 versus 26% for 200 ng BSA). However, the digestion time was shortened from 12 h to 50 s. When the enzymatic activity was decreased after seven runs, the IMER could be easily regenerated by removing Cu(2+) via EDTA followed by trypsin immobilization with fresh Cu(2+) introduced, yielding the equal sequence coverage (26% for 200 ng BSA). For similar to 5 mu g rat liver extract, even more proteins were identified with the immobilized trypsin digestion within 150 s in comparison to the in-solution digestion for 24 h (541 versus 483), demonstrating that the IMER could be a promising tool for efficient and high-throughput proteome profiling.
Language英语
WOS IDWOS:000288137300015
Citation statistics
Cited Times:37[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/115314
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang LH(张丽华)
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
Recommended Citation
GB/T 7714
Ma, Junfeng,Hou, Chunyan,Liang, Yu,et al. Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith[J]. PROTEOMICS,2011,11(5):991-995.
APA Ma, Junfeng.,Hou, Chunyan.,Liang, Yu.,Wang, Tingting.,Liang, Zhen.,...&张丽华.(2011).Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith.PROTEOMICS,11(5),991-995.
MLA Ma, Junfeng,et al."Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith".PROTEOMICS 11.5(2011):991-995.
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