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学科主题物理化学
Hydrophilic immobilized trypsin reactor with magnetic graphene oxide as support for high efficient proteome digestion
Jiang, Bo1,2,3; Yang, Kaiguang1; Zhao, Qun1,2; Wu, Qi1,2; Liang, Zhen1; Zhang, Lihua1; Peng, Xiaojun3; Zhang, Yukui1; Zhang LH(张丽华)
关键词Immobilized Enzymatic Reactor Graphene Oxide Magnetic Nanoparticles Proteomics
刊名JOURNAL OF CHROMATOGRAPHY A
2012-09-07
DOI10.1016/j.chroma.2012.07.030
1254页:8-13
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 ; Life Sciences & Biomedicine ; Physical Sciences
类目[WOS]Biochemical Research Methods ; Chemistry, Analytical
研究领域[WOS]Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]ON-PLATE PROTEOLYSIS ; MASS-SPECTROMETRY ; MS ANALYSIS ; SILICA ; NANOPARTICLES ; ONLINE ; MICROREACTOR ; FABRICATION ; MONOLITHS ; SHEETS
英文摘要In this paper, magnetic Fe3O4 nanoparticles modified graphene oxide nanocomposites (GO-CO-NH-Fe3O4) were prepared by covalent bonding, via the reaction between the amino groups of fuctionalized Fe3O4 and the carboxylic groups of GO, confirmed by Fourier-transform infrared spectra, Raman spectroscopy, and transmission electron microscopy. With GO-CO-NH-Fe3O4 as a novel substrate, trypsin was immobilized via pi-pi stacking and hydrogen bonding interaction, and the binding capacity of trypsin reached as high as 0.275 mg/mg. Since GO-CO-NH-Fe3O4 worked as not only support for enzyme immobilization, but also as an excellent microwave irradiation absorber, the digestion efficiency could be further improved with microwave assistance. By such an immobilized enzymatic reactor (IMER), standard proteins could be efficiently digested within 15 s, with sequence coverages comparable or better than those obtained by conventional in-solution digestion (12 h). Since trypsin was immobilized under mild conditions, the enzymatic activity of IMER preserved at least for a month. In addition, due to the good hydrophilicity of GO, no peptide residue was observed in the sequent digestion of bovine serum albumin and myoglobin. To further confirm the efficiency of such an IMER for proteome analysis, it was applied to digest proteins extracted from rat liver, followed by nanoRPLC-ESI-MS/MS analysis. With only 5 min microwave-assisted digestion, in 3 parallel runs, totally 456 protein groups were identified, comparable to that obtained by 12 h in-solution digestion, indicating the great potential of IMERs with GO-CO-NH-Fe3O4 as the support for high throughput proteome study. (c) 2012 Elsevier B.V. All rights reserved.
语种英语
WOS记录号WOS:000308048000002
引用统计
被引频次:61[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/321008/117993
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, Key Lab Separat Sci Analyt Chem, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
3.Dalian Univ Technol, State Key Lab Fine Chem, Dalian 116012, Peoples R China
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GB/T 7714
Jiang, Bo,Yang, Kaiguang,Zhao, Qun,et al. Hydrophilic immobilized trypsin reactor with magnetic graphene oxide as support for high efficient proteome digestion[J]. JOURNAL OF CHROMATOGRAPHY A,2012,1254:8-13.
APA Jiang, Bo.,Yang, Kaiguang.,Zhao, Qun.,Wu, Qi.,Liang, Zhen.,...&张丽华.(2012).Hydrophilic immobilized trypsin reactor with magnetic graphene oxide as support for high efficient proteome digestion.JOURNAL OF CHROMATOGRAPHY A,1254,8-13.
MLA Jiang, Bo,et al."Hydrophilic immobilized trypsin reactor with magnetic graphene oxide as support for high efficient proteome digestion".JOURNAL OF CHROMATOGRAPHY A 1254(2012):8-13.
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