期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1216, 期 28, 页码 5377-5384出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2009.05.030
关键词
Peptidomics; Restricted access material (RAM); Multidimensional separation; Serum peptides; Proteomics
资金
- National Natural Sciences Foundation of China [20675081, 20735004, 20605022]
- China State Key Basic Research Program [2005CB522701, 2007CB914104]
- China High Technology Research Program [2006AA02A309]
- Knowledge Innovation program of CAS [KJCX2.YW.HO9]
- Knowledge Innovation Program of DICP
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.
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