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题名: 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
作者: Hu, Lianghai1;  Boos, Karl-Siegfried2;  Ye, Mingliang1;  Wu, Ren'an1;  Zou, Hanfa1
关键词: Peptidomics ;  Restricted access material (RAM) ;  Multidimensional separation ;  Serum peptides ;  Proteomics
刊名: JOURNAL OF CHROMATOGRAPHY A
发表日期: 2009-07-10
DOI: 10.1016/j.chroma.2009.05.030
卷: 1216, 期:28, 页:5377-5384
收录类别: SCI
文章类型: Article
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
英文摘要: As 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.
关键词[WOS]: PERFORMANCE LIQUID-CHROMATOGRAPHY ;  SOLID-PHASE EXTRACTION ;  BIOMARKER DISCOVERY ;  SAMPLE PREPARATION ;  PROTEOME ANALYSIS ;  SILICA PARTICLES ;  YEAST PROTEOME ;  HUMAN PLASMA ;  MS/MS ;  IDENTIFICATION
语种: 英语
WOS记录号: WOS:000267557800006
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/141606
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.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:
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.
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