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Fast on-column protein digestion with subsequent peptide mapping using tandem mass spectrometry with information dependent acquisition

Hedström, Martin LU ; Andersson, Maria LU ; Galaev, Igor LU and Mattiasson, Bo LU (2005) In Journal of Chromatography A 1080(2). p.117-123
Abstract
A platform for rapid on-line protein digestion of protein mixtures for direct infusion to a mass spectrometer is presented. A mixture of protein A, staphylococcal enterotoxin B and cytochrome c was used as a model mixture injected on a gel filtration column and a trypsin reactor which were connected in series to a micro liquid chromatography (mu LC) system. The peptides in the column eluate were analyzed with ESI tandem mass spectrometry, utilizing information dependent acquisition (IDA). In one step, the proteins in the mixture (mu M concentrations) were concomitantly desalted, separated, digested and identified with an overall analysis time of less than 40 min. Protein sequence coverage of 78-95% for the involved substances was achieved.... (More)
A platform for rapid on-line protein digestion of protein mixtures for direct infusion to a mass spectrometer is presented. A mixture of protein A, staphylococcal enterotoxin B and cytochrome c was used as a model mixture injected on a gel filtration column and a trypsin reactor which were connected in series to a micro liquid chromatography (mu LC) system. The peptides in the column eluate were analyzed with ESI tandem mass spectrometry, utilizing information dependent acquisition (IDA). In one step, the proteins in the mixture (mu M concentrations) were concomitantly desalted, separated, digested and identified with an overall analysis time of less than 40 min. Protein sequence coverage of 78-95% for the involved substances was achieved. (c) 2005 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Chromatography A
volume
1080
issue
2
pages
117 - 123
publisher
Elsevier
external identifiers
  • wos:000230238600003
  • pmid:16008049
  • scopus:20444484810
ISSN
0021-9673
DOI
10.1016/j.chroma.2005.04.069
language
English
LU publication?
yes
id
3738f58f-c701-41d2-b1c0-1c7282f3ac63 (old id 155109)
date added to LUP
2007-07-02 11:30:15
date last changed
2017-01-01 06:47:03
@article{3738f58f-c701-41d2-b1c0-1c7282f3ac63,
  abstract     = {A platform for rapid on-line protein digestion of protein mixtures for direct infusion to a mass spectrometer is presented. A mixture of protein A, staphylococcal enterotoxin B and cytochrome c was used as a model mixture injected on a gel filtration column and a trypsin reactor which were connected in series to a micro liquid chromatography (mu LC) system. The peptides in the column eluate were analyzed with ESI tandem mass spectrometry, utilizing information dependent acquisition (IDA). In one step, the proteins in the mixture (mu M concentrations) were concomitantly desalted, separated, digested and identified with an overall analysis time of less than 40 min. Protein sequence coverage of 78-95% for the involved substances was achieved. (c) 2005 Elsevier B.V. All rights reserved.},
  author       = {Hedström, Martin and Andersson, Maria and Galaev, Igor and Mattiasson, Bo},
  issn         = {0021-9673},
  language     = {eng},
  number       = {2},
  pages        = {117--123},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Fast on-column protein digestion with subsequent peptide mapping using tandem mass spectrometry with information dependent acquisition},
  url          = {http://dx.doi.org/10.1016/j.chroma.2005.04.069},
  volume       = {1080},
  year         = {2005},
}