DICP OpenIR
Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS
Duan, JC; Liang, Z; Yang, C; Zhang, J; Zhang, LH; Zhang, WB; Zhang, YK; Zhang YK(张玉奎); Zhang YK(张玉奎)
KeywordEnzymatic Reactor Lc-ms/ms Monolithic Column Protein Identification
Source PublicationPROTEOMICS
2006
DOI10.1002/pmic.200500234
Volume6Issue:2Pages:412-419
Indexed ByISTP ; SCI
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-ELECTROPHORESIS ; AFFINITY-CHROMATOGRAPHY ; PROTEOLYTIC DIGESTION ; TRYPSIN MICROREACTOR ; PEPTIDE SEPARATION ; ION-EXCHANGE ; SOL-GEL ; PERFORMANCE ; CHIP
AbstractA monolithic enzymatic microreactor was prepared in a fused-silica capillary by in situ polymerization of acrylamide, glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) in the presence of a binary porogenic mixture of dodecanol and cyclohexanol, followed by ammonia solution treatment, glutaraldehyde activation and trypsin modification. The choice of acrylamide as co-monomer was found useful to improve the efficiency of trypsin modification, thus, to increase the enzyme activity. The optimized microreactor offered very low back pressure, enabling the fast digestion of proteins flowing through the reactor. The performance of the monolithic microreactor was demonstrated with the digestion of cytochrome c at high flow rate. The digests were then characterized by CE and HPLC-MS/MS with the sequence coverage of 57.7%. The digestion efficiency was found over 230 times as high as that of the conventional method. in addition, for the first time, protein digestion carried out in a mixture of water and ACN was compared with the conventional aqueous reaction using MS/MS detection, and the former solution was found more compatible and more efficient for protein digestion.
Language英语
URL查看原文
WOS IDWOS:000235207700003
Citation statistics
Cited Times:77[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/93527
Collection中国科学院大连化学物理研究所
Corresponding AuthorZhang YK(张玉奎); Zhang YK(张玉奎)
AffiliationChinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China
Recommended Citation
GB/T 7714
Duan, JC,Liang, Z,Yang, C,et al. Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS[J]. PROTEOMICS,2006,6(2):412-419.
APA Duan, JC.,Liang, Z.,Yang, C.,Zhang, J.,Zhang, LH.,...&张玉奎.(2006).Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS.PROTEOMICS,6(2),412-419.
MLA Duan, JC,et al."Rapid protein identification using monolithic enzymatic microreactor and LC-ESI-MS/MS".PROTEOMICS 6.2(2006):412-419.
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