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
Abstract
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 microL 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 microL human serum sample.
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SPEs can last from several hours to several weeks.
“SPEs can last from several hours to several weeks [29], [30], [31].”
From Hindlimb suspension and spe-like radiation impairs clearance of bacterial infections (2014) · cites this paper as [30]
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- Authors
- Hu, L., Boos, K.-S., Ye, M., Wu, R., & Zou, H.
- Affiliations
- Dalian Institute of Chemical Physics (CN)
- Ludwig-Maximilians-Universität München (DE)
- Corresponding author
- Zou, Hanfa
- Venue
- Journal of Chromatography A
- Publisher
- Elsevier BV
- Published
- 2009-07
- Volume
- 1216
- Issue
- 28
- Pages
- 5377-5384
- ISSN
- 0021-9673 (ISSN-L)
- OpenAlex
- W2075173429
- Cited by
- 56
- References
- 31
- Citations by year
- 2022: 4 · 2021: 1 · 2019: 3 · 2018: 2 · 2017: 3 · 2015: 5 · 2014: 10 · 2013: 5 · 2012: 7
- FWCI
- 3.29
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Chromatography A (publishedVersion)
- PubMed (publishedVersion)
- Dalian Institute of Chemical Physics, CAS Institutional Repository (submittedVersion)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Keywords
- serum peptidome, restricted access materials, size exclusion chromatography, multidimensional separation, human serum, biomarker discovery, sulfonic acid functionalization
- MeSH terms
- Chromatography, Liquid, Equipment Design, Humans, Molecular Weight, Peptides, Reproducibility of Results, Proteomics, Solid Phase Extraction, Tandem Mass Spectrometry
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