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Automated multi-dimensional liquid chromatography: sample preparation and identification of peptides from human blood filtrate

Machtejevas, E; John, H; Wagner, K; Standker, L; Marko-Varga, György LU ; Forssmann, WG; Bischoff, R and Unger, KK (2004) In Journal of Chromatography. B 803(1). p.121-130
Abstract
A comprehensive on-line sample clean-up with an integrated two-dimensional HPLC system was developed for the analysis of natural peptides. Samples comprised of endogenous peptides with molecular weights up to 20 kDa were generated from human hemofiltrate (HF) obtained from patients with chronic renal failure. The (poly-)peptides were separated using novel silica-based restricted access materials with strong cation-exchange functionalities (SCX-RAM). The size-selective sample fractionation step is followed by cation-exchange chromatography as the first dimension. The subsequent second dimension of separation is based on hydrophobic interaction using four parallel short reversed-phase (RP) columns implemented via a fully automated column... (More)
A comprehensive on-line sample clean-up with an integrated two-dimensional HPLC system was developed for the analysis of natural peptides. Samples comprised of endogenous peptides with molecular weights up to 20 kDa were generated from human hemofiltrate (HF) obtained from patients with chronic renal failure. The (poly-)peptides were separated using novel silica-based restricted access materials with strong cation-exchange functionalities (SCX-RAM). The size-selective sample fractionation step is followed by cation-exchange chromatography as the first dimension. The subsequent second dimension of separation is based on hydrophobic interaction using four parallel short reversed-phase (RP) columns implemented via a fully automated column switching technique. More than 1000 peaks were resolved within the total analysis time of 96 min. Substances of selected peaks were sampled to analyse their molecular weights by off-line MALDI-TOF mass spectrometry and to determine their amino acid sequence by Edman degradation. The potential for comprehensive peptide mapping and identification is demonstrated. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
sample preparation, identification, human blood filtrate, peptides
in
Journal of Chromatography. B
volume
803
issue
1
pages
121 - 130
publisher
Elsevier
external identifiers
  • wos:000220343500013
  • pmid:15026005
  • scopus:1542719143
ISSN
1873-376X
DOI
10.1016/j.jchromb.2003.07.015
language
English
LU publication?
yes
id
58bb7464-4b65-4461-983c-8540a1e4aa6a (old id 284592)
date added to LUP
2007-08-03 12:30:01
date last changed
2017-08-20 03:40:57
@article{58bb7464-4b65-4461-983c-8540a1e4aa6a,
  abstract     = {A comprehensive on-line sample clean-up with an integrated two-dimensional HPLC system was developed for the analysis of natural peptides. Samples comprised of endogenous peptides with molecular weights up to 20 kDa were generated from human hemofiltrate (HF) obtained from patients with chronic renal failure. The (poly-)peptides were separated using novel silica-based restricted access materials with strong cation-exchange functionalities (SCX-RAM). The size-selective sample fractionation step is followed by cation-exchange chromatography as the first dimension. The subsequent second dimension of separation is based on hydrophobic interaction using four parallel short reversed-phase (RP) columns implemented via a fully automated column switching technique. More than 1000 peaks were resolved within the total analysis time of 96 min. Substances of selected peaks were sampled to analyse their molecular weights by off-line MALDI-TOF mass spectrometry and to determine their amino acid sequence by Edman degradation. The potential for comprehensive peptide mapping and identification is demonstrated.},
  author       = {Machtejevas, E and John, H and Wagner, K and Standker, L and Marko-Varga, György and Forssmann, WG and Bischoff, R and Unger, KK},
  issn         = {1873-376X},
  keyword      = {sample preparation,identification,human blood filtrate,peptides},
  language     = {eng},
  number       = {1},
  pages        = {121--130},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography. B},
  title        = {Automated multi-dimensional liquid chromatography: sample preparation and identification of peptides from human blood filtrate},
  url          = {http://dx.doi.org/10.1016/j.jchromb.2003.07.015},
  volume       = {803},
  year         = {2004},
}