DICP OpenIR
Subject Area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)
KeywordImmobilized Enzymatic Reactor Graphene Oxide Magnetic Nanoparticles Proteomics
Source PublicationJOURNAL OF CHROMATOGRAPHY A
2012-09-07
DOI10.1016/j.chroma.2012.07.030
Volume1254Pages:8-13
Indexed BySCI
SubtypeArticle
Department18
Funding Project1810
Contribution Rank1,1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordON-PLATE PROTEOLYSIS ; MASS-SPECTROMETRY ; MS ANALYSIS ; SILICA ; NANOPARTICLES ; ONLINE ; MICROREACTOR ; FABRICATION ; MONOLITHS ; SHEETS
AbstractIn 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.
Language英语
WOS IDWOS:000308048000002
Citation statistics
Cited Times:71[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/117993
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, 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
Recommended Citation
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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