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Differential proteomic analysis of HT29 Cl.16E cells line and intestinal epithelial cells by LC ESI/QTOF mass spectrometry.

Nanni, P ; Mezzanottea, L ; Roda, G ; Caponi, A ; Levander, Fredrik LU ; James, Peter LU orcid and Roda, A (2009) In Journal of Proteomics 72(5). p.865-873
Abstract
Abstract in Undetermined
Intestinal epithelial cells (IECs) play a key role in Crohn's disease, a chronic inflammatory bowel disease which requires invasive examinations to be diagnosed. The comparison of the cellular protein expression profiles of Crohn's disease patients and healthy subjects is fundamental for the identification of proteins clinically relevant as new biomarkers or as drug targets. For this purpose a differential label-free nano-LC ESI/QTOF mass spectrometry (MS) approach combined with targeted MS/MS analysis has been developed and applied to isolated IECs.

We report here a study of the protein variations in IECs from healthy subjects (H) and Crohn's disease patients (CD). The method was previously validated... (More)
Abstract in Undetermined
Intestinal epithelial cells (IECs) play a key role in Crohn's disease, a chronic inflammatory bowel disease which requires invasive examinations to be diagnosed. The comparison of the cellular protein expression profiles of Crohn's disease patients and healthy subjects is fundamental for the identification of proteins clinically relevant as new biomarkers or as drug targets. For this purpose a differential label-free nano-LC ESI/QTOF mass spectrometry (MS) approach combined with targeted MS/MS analysis has been developed and applied to isolated IECs.

We report here a study of the protein variations in IECs from healthy subjects (H) and Crohn's disease patients (CD). The method was previously validated using HT29 Cl.16E cell line, normal or treated with interferon-gamma as a model of inflammation.

Subcellular fractions proteins were extracted from HT29 and IECs and for each fraction monodimensional gel-electrophoresis was performed and the proteins subjected to tryptic digestion. The resulting peptides were analysed by LC ESI/QTOF MS and the obtained chromatographic runs were aligned with msInspect software. The peptides differently expressed were statistically evaluated using the Proteios Software Environment (ProSE) and identified by LC ESI/QTOF MS/MS analysis and database search.

The preliminary results obtained allowed the identification of many proteins involved in the inflammation processes. (Less)
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author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Crohn's disease, HT29 Cl.16E, Intestinal epithelial cells, Label-free mass spectrometry, Proteomics
in
Journal of Proteomics
volume
72
issue
5
pages
865 - 873
publisher
Elsevier
external identifiers
  • wos:000268604400015
  • scopus:67649531403
ISSN
1874-3919
DOI
10.1016/j.jprot.2008.12.010
language
English
LU publication?
yes
id
4f284fb2-6f3b-4e84-8536-6680e6431a98 (old id 1288176)
date added to LUP
2016-04-01 11:41:33
date last changed
2023-09-01 03:32:38
@article{4f284fb2-6f3b-4e84-8536-6680e6431a98,
  abstract     = {{Abstract in Undetermined<br/>Intestinal epithelial cells (IECs) play a key role in Crohn's disease, a chronic inflammatory bowel disease which requires invasive examinations to be diagnosed. The comparison of the cellular protein expression profiles of Crohn's disease patients and healthy subjects is fundamental for the identification of proteins clinically relevant as new biomarkers or as drug targets. For this purpose a differential label-free nano-LC ESI/QTOF mass spectrometry (MS) approach combined with targeted MS/MS analysis has been developed and applied to isolated IECs.<br/><br/>We report here a study of the protein variations in IECs from healthy subjects (H) and Crohn's disease patients (CD). The method was previously validated using HT29 Cl.16E cell line, normal or treated with interferon-gamma as a model of inflammation.<br/><br/>Subcellular fractions proteins were extracted from HT29 and IECs and for each fraction monodimensional gel-electrophoresis was performed and the proteins subjected to tryptic digestion. The resulting peptides were analysed by LC ESI/QTOF MS and the obtained chromatographic runs were aligned with msInspect software. The peptides differently expressed were statistically evaluated using the Proteios Software Environment (ProSE) and identified by LC ESI/QTOF MS/MS analysis and database search.<br/><br/>The preliminary results obtained allowed the identification of many proteins involved in the inflammation processes.}},
  author       = {{Nanni, P and Mezzanottea, L and Roda, G and Caponi, A and Levander, Fredrik and James, Peter and Roda, A}},
  issn         = {{1874-3919}},
  keywords     = {{Crohn's disease; HT29 Cl.16E; Intestinal epithelial cells; Label-free mass spectrometry; Proteomics}},
  language     = {{eng}},
  number       = {{5}},
  pages        = {{865--873}},
  publisher    = {{Elsevier}},
  series       = {{Journal of Proteomics}},
  title        = {{Differential proteomic analysis of HT29 Cl.16E cells line and intestinal epithelial cells by LC ESI/QTOF mass spectrometry.}},
  url          = {{http://dx.doi.org/10.1016/j.jprot.2008.12.010}},
  doi          = {{10.1016/j.jprot.2008.12.010}},
  volume       = {{72}},
  year         = {{2009}},
}