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学科主题物理化学
High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor
Wu, Shuaibin1,2; Sun, Liangliang1,2; Ma, Junfeng1,2; Yang, Kaiguang1; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
关键词Immobilized Enzyme Reactor Poly (Acrylamide-co-methylenebisacrylamide) Monolith Protein Digestion Protein Identification Proteomics
刊名TALANTA
2011-02-15
ISSN待补充
DOI10.1016/j.talanta.2010.12.011
83期:5页:1748-1753
收录类别SCI
文章类型Article
部门归属18
项目归属1810
产权排名1,1
WOS标题词Science & Technology ; Physical Sciences
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]ENABLING PROTEIN DIGESTION ; MASS-SPECTROMETRY ; CAPILLARY ELECTROCHROMATOGRAPHY ; PEPTIDE SEPARATION ; TRYPSIN REACTOR ; IDENTIFICATION ; PROTEOMICS ; SYSTEM ; ONLINE
英文摘要High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor; A poly (acrylamide-co-methylenebisacrylamide) (poly (AAm-co-MBA)) monolith was prepared by thermal polymerization in the 100 or 250 mu m i.d. capillary. The monolithic support was activated by ethylenediamine followed by glutaraldehyde. Trypsin was then introduced to form an immobilized enzyme reactor (IMER). The prepared IMER showed a reliable mechanical stability and permeability (permeability constant K = 2.65 x 10(-13) m(2)). With BSA as the model protein, efficient digestion was completed within 20 s, yielding the sequence coverage of 57%, better than that obtained from the traditional in-solution digestion (42%), which took about 12 h. Moreover. BSA down to femtomole was efficiently digested by the IMER and positively identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). To test the applicability of IMER for complex sample profiling, proteins extracted from Escherichia coli were digested by the IMER and further analyzed by nanoreversed phase liquid chromatography-electrospray ionization-mass spectrometry (nanoRPLC-ESI-MS/MS). In comparison to in-solution digestion, despite slightly fewer proteins were positively identified at a false discovery rate (FDR) of similar to 1% (333 vs 411), the digestion time used was largely shortened (20 s vs 24 h), implying superior digestion performance for the high throughput analysis of complex samples. (C) 2011 Elsevier B.V. All rights reserved.
语种英语
WOS记录号WOS:000287110100065
引用统计
被引频次:34[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/115315
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
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Wu, Shuaibin,Sun, Liangliang,Ma, Junfeng,et al. High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor[J]. TALANTA,2011,83(5):1748-1753.
APA Wu, Shuaibin.,Sun, Liangliang.,Ma, Junfeng.,Yang, Kaiguang.,Liang, Zhen.,...&张丽华.(2011).High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor.TALANTA,83(5),1748-1753.
MLA Wu, Shuaibin,et al."High throughput tryptic digestion via poly (acrylamide-co-methylenebisacrylamide) monolith based immobilized enzyme reactor".TALANTA 83.5(2011):1748-1753.
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