Format

Send to

Choose Destination
J Biomed Biotechnol. 2010;2010:927917. doi: 10.1155/2010/927917. Epub 2009 Nov 1.

A proteomic approach for plasma biomarker discovery with iTRAQ labelling and OFFGEL fractionation.

Author information

1
Centre INSERM Régional de Recherche sur le Cancer U892, Centre Régional de Lutte Contre le Cancer Paul Papin, 2 rue Moll, 49933 Angers Cedex 9, France.

Abstract

Human blood plasma contains a plethora of proteins, encompassing not only proteins that have plasma-based functionalities, but also possibly every other form of low concentrated human proteins. As it circulates through the tissues, the plasma picks up proteins that are released from their origin due to physiological events such as tissue remodeling and cell death. Specific disease processes or tumors are often characterized by plasma "signatures," which may become obvious via changes in the plasma proteome profile, for example, through over expression of proteins. However, the wide dynamic range of proteins present in plasma makes their analysis very challenging, because high-abundance proteins tend to mask those of lower abundance. In the present study, we used a strategy combining iTRAQ as a reagent which improved the peptide ionization and peptide OFFGEL fractionation that has already been shown, in our previous research, to improve the proteome coverage of cellular extracts. Two prefractioning methods were compared: immunodepletion and a bead-based library of combinatorial hexapeptide technology. Our data suggested that both methods were complementary, with regard to the number of identified proteins. iTRAQ labelling, in association with OFFGEL fractionation, allowed more than 300 different proteins to be characterized from 400 microg of plasma proteins.

PMID:
19888438
PMCID:
PMC2771280
DOI:
10.1155/2010/927917
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Hindawi Limited Icon for PubMed Central
Loading ...
Support Center