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Selective on-line serum peptide extraction and multidimensional separation by coupling a restricted-access material-based capillary trap column with nanoliquid chromatography-tandem mass spectrometry
Hu, Lianghai1; Boos, Karl-Siegfried2; Ye, Mingliang1; Wu, Ren'an1; Zou, Hanfa1
KeywordPeptidomics Restricted Access Material (Ram) Multidimensional Separation Serum Peptides Proteomics
Source PublicationJOURNAL OF CHROMATOGRAPHY A
2009-07-10
DOI10.1016/j.chroma.2009.05.030
Volume1216Issue:28Pages:5377-5384
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS KeywordPERFORMANCE LIQUID-CHROMATOGRAPHY ; SOLID-PHASE EXTRACTION ; BIOMARKER DISCOVERY ; SAMPLE PREPARATION ; PROTEOME ANALYSIS ; SILICA PARTICLES ; YEAST PROTEOME ; HUMAN PLASMA ; MS/MS ; IDENTIFICATION
AbstractAs the serum peptidome gets increasing attention for biomarker discovery, one of the important issues is how to efficiently extract the peptides from highly complex human serum for peptidome analysis. Here we developed a fully automated platform for direct injection, on-line extraction, multidimensional separation and MS detection of peptides present in human serum. A capillary SPE column packed with a novel mix mode restricted access material (RAM) exhibiting strong cation exchange and size exclusion chromatography (SCX/SEC) properties were coupled with a nanoliquid chromatography-mass spectrometry (nanoLC-MS) system. The capillary SPE column excludes the high abundant serum proteins such as HSA by size exclusion chromatography and simultaneously extracts the low molecular weight peptides by binding to sulfonic acid residues. Subsequently, the trapped peptides are eluted to a capillary LC column packed with a RP-C18 stationary phase. After injection of only 2 mu L human serum to the one-dimensional nanoLC-MS system around 400 peptides could be identified. When conducting a multidimensional separation, the described SCX/SEC/RP-MS platform allows the separation and identification of 1286 peptides present in human serum by the injection and on-line processing of 20 mu L human serum sample. (C) 2009 Elsevier B.V. All rights reserved.
Language英语
WOS IDWOS:000267557800006
Citation statistics
Cited Times:43[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/141606
Collection中国科学院大连化学物理研究所
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.Univ Munich, Med Ctr, Inst Clin Chem, Lab BioSeparat, D-81366 Munich, Germany
Recommended Citation
GB/T 7714
Hu, Lianghai,Boos, Karl-Siegfried,Ye, Mingliang,et al. Selective on-line serum peptide extraction and multidimensional separation by coupling a restricted-access material-based capillary trap column with nanoliquid chromatography-tandem mass spectrometry[J]. JOURNAL OF CHROMATOGRAPHY A,2009,1216(28):5377-5384.
APA Hu, Lianghai,Boos, Karl-Siegfried,Ye, Mingliang,Wu, Ren'an,&Zou, Hanfa.(2009).Selective on-line serum peptide extraction and multidimensional separation by coupling a restricted-access material-based capillary trap column with nanoliquid chromatography-tandem mass spectrometry.JOURNAL OF CHROMATOGRAPHY A,1216(28),5377-5384.
MLA Hu, Lianghai,et al."Selective on-line serum peptide extraction and multidimensional separation by coupling a restricted-access material-based capillary trap column with nanoliquid chromatography-tandem mass spectrometry".JOURNAL OF CHROMATOGRAPHY A 1216.28(2009):5377-5384.
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