title: Evaluation of isotope-coded protein labeling (ICPL) in the quantitative analysis of complex proteomes.
abstract: 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.
authors.affiliation: Laboratorio de Proteómica, Centro Nacional de Biotecnología, Consejo Superior de Investigaciones Científicas (CSIC), Madrid, Spain. alberto.paradela@cnb.csic.es
authors.forename: Alberto
authors.initials: A
authors.lastname: Paradela
authors.forename: Miguel
authors.initials: M
authors.lastname: Marcilla
authors.forename: Rosana
authors.initials: R
authors.lastname: Navajas
authors.forename: Laura
authors.initials: L
authors.lastname: Ferreira
authors.forename: Antonio
authors.initials: A
authors.lastname: Ramos-Fernandez
authors.forename: Marisol
authors.initials: M
authors.lastname: Fernández
authors.forename: Javier Fernando
authors.initials: JF
authors.lastname: Mariscotti
authors.forename: Francisco
authors.initials: F
authors.lastname: García-del Portillo
authors.forename: Juan Pablo
authors.initials: JP
authors.lastname: Albar
mesh_terms: D002247:Carbon Isotopes / Q000737:chemistry; D019411:Clinical Laboratory Techniques; D019295:Computational Biology; D015180:Electrophoresis, Gel, Two-Dimensional; D007553:Isotope Labeling / Q000379:methods*; D009587:Nitrogen Isotopes / Q000737:chemistry; D011506:Proteins / Q000032:analysis; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*
chemical_list: D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D020543:Proteome