DICP OpenIR
Subject Area物理化学
Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics
Wu, Shuaibin1,3; Zhang, Lei; Yang, Kaiguang1; Liang, Zhen1; Zhang, Lihua; Zhang, Yukui1; Yang KG(杨开广)
KeywordMetal-ion Chelation Polyacrylamide Monolith Polyethylenimine Tryptic Digestion Regenerable Immobilized Enzyme Reactor
Source PublicationANALYTICAL AND BIOANALYTICAL CHEMISTRY
2012
DOI10.1007/s00216-011-5501-6
Volume402Issue:2Pages:703-710
Indexed BySCI
SubtypeArticle
Department18
Funding Project1810
Contribution Rank1,1
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordLASER-DESORPTION/IONIZATION-TIME ; MAGNETIC SILICA MICROSPHERES ; PHASE LIQUID-CHROMATOGRAPHY ; EFFICIENT PROTEIN DIGESTION ; TANDEM MASS-SPECTROMETRY ; TRYPSIN IMMOBILIZATION ; IDENTIFICATION ; ONLINE ; MICROREACTOR ; ADSORPTION
AbstractInitially, a poly (glycidyl methacrylate-co-acrylamide-co-methylenebisacrylamide) monolith was prepared in the 100 mu m i.d. capillary, and then was grafted with polyethylenimine (Mw, similar to 25,000) for adsorbing Cu(2+), followed by chelating trypsin. As a result, efficient digestion for BSA (100 ng/mu L) was completed within 50 s via such immobilized enzyme reactor (IMER); yielding 47% sequence coverage by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis. Compared with the conventional method for preparing the metal-ion chelated IMER, the regeneration of such IMER can be achieved facilely by the respective 30 min desorption and re-adsorption of trypsin, and 51% sequence coverage was obtained for 50 s BSA digestion after regeneration. BSA down to femtomole was also efficiently digested by the prepared regenerable IMER. Meanwhile, after the consecutive digestion of myoglobin and BSA, there was not any mutual interference for both during MALDI-TOF MS identification, indicating the low nonspecific adsorption of such regenerable IMER. To test the applicability of regenerable IMER for complex sample profiling, proteins (150 ng) extracted from Escherichia coli were digested within 80 s by the regenerable IMER and further analyzed by nanoreversed phase liquid chromatography-electrospray ionization-mass spectrometry successfully, showing its practicability for the high throughput analysis of complex samples.
Language英语
WOS IDWOS:000298645300012
Citation statistics
Cited Times:22[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/118276
Collection中国科学院大连化学物理研究所
Corresponding AuthorYang KG(杨开广)
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Dalian Inst Chem Phys, State Key Lab Catalysis, Dalian 116023, Peoples R China
3.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Wu, Shuaibin,Zhang, Lei,Yang, Kaiguang,et al. Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics[J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY,2012,402(2):703-710.
APA Wu, Shuaibin.,Zhang, Lei.,Yang, Kaiguang.,Liang, Zhen.,Zhang, Lihua.,...&杨开广.(2012).Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics.ANALYTICAL AND BIOANALYTICAL CHEMISTRY,402(2),703-710.
MLA Wu, Shuaibin,et al."Preparing a metal-ion chelated immobilized enzyme reactor based on the polyacrylamide monolith grafted with polyethylenimine for a facile regeneration and high throughput tryptic digestion in proteomics".ANALYTICAL AND BIOANALYTICAL CHEMISTRY 402.2(2012):703-710.
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