Evaluation of isotope-coded protein labeling (ICPL) in the quantitative analysis of complex proteomes

An evaluation of the ICPL (isotope-coded protein labeling) non-isobaric labeling technique was performed using two different biological models. Two samples containing phage T4 capsids were mixed in a 1:1 ratio after being labeled with the light or heavy versions of the ICPL reagent. The analysis of...

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Bibliographic Details
Published in:Talanta (Oxford) 2010-02, Vol.80 (4), p.1496-1502
Main Authors: Paradela, Alberto, Marcilla, Miguel, Navajas, Rosana, Ferreira, Laura, Ramos-Fernandez, Antonio, Fernández, Marisol, Mariscotti, Javier Fernando, Portillo, Francisco García-del, Albar, Juan Pablo
Format: Article
Language:English
Subjects:
Carbon Isotopes - chemistry
Clinical Laboratory Techniques
Computational Biology
Electrophoresis, Gel, Two-Dimensional
ICPL
Isotope Labeling - methods
Mass spectrometry
Nitrogen Isotopes - chemistry
Phage T4
Proteins - analysis
Proteome - analysis
Proteomics - methods
Quantitative proteomics
Salmonella enterica
Stable isotopic labeling
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Summary:An evaluation of the ICPL (isotope-coded protein labeling) non-isobaric labeling technique was performed using two different biological models. Two samples containing phage T4 capsids were mixed in a 1:1 ratio after being labeled with the light or heavy versions of the ICPL reagent. The analysis of this proteome demonstrated the feasibility of this approach for differential quantitative proteomics and was employed to optimize the experimental parameters of the ICPL workflow. ICPL-mediated analysis of two more complex proteomes, those of a Salmonella enterica serovar Typhimurium virulent strain and an isogenic attenuated mutant, and its comparison with the results obtained in a 2D-PAGE “classical” approach confirmed that ICPL is a valuable alternative to other labeling techniques currently in use. In addition, our results suggest that labeling at the peptide level instead of following the standard ICPL workflow should increase both the number of proteins quantified and the reliability of the quantification.
ISSN:0039-9140
1873-3573
DOI:10.1016/j.talanta.2009.06.083