[Introduction and Objectives] Peripheral blood mononuclear cells (PBMCs) are a popular model system to study the physiological and metabolic activity of cells within the body. PBMCs have enabled a very broad collection of biomedical applications. Monitoring gene expression and posttranslational modifications are very promising areas in biomarker discovery and translational research. In this study, we aimed to have the most extensive proteome map of PBMCs and monitor the in vitro effect of reactive peroxide at low concentration. [Methods] PBMCs from a healthy male individual were purchased from AllCells. 1mill cells aliquots were in vitro treated with 5 mM H2O2 for 0, 2, 10, 30 and 80 min. Sample preparation was performed using the Mass Spec Sample Prep Kit for Cultured Cells (Pierce, Rockford IL). Peptide digests were then analyzed by LC-MS/MS analysis on a Thermo ScientificTM Q ExactiveTM HF mass spectrometer. Database search and oxidation site localization were performed using Thermo ScientificTM Protein Discoverer 2.0. Inferno was then used for further statistical analysis and ProteinCenter was used to extract biological context. Results and Discussion: Access to the complete atlas of gene expressions and posttranslational modifications in PBMCs will permit more sophisticated studies such us the selection of potential biomarkers that could be used for many purposes ranging from diagnostic, prognosis or even help selecting the appropriate therapy for a patient. The bioinformatic analysis of the results yielded the identification of over 8000 proteins. In addition, over 5000 proteins were accurately quantified and over 7000 oxidation events were identified. [Conclusion] Overall, this study not only adds significant value in the mechanistic understanding of redox signalling, but it also creates a valuable protein repository that could lead to the development of new therapeutic strategies. This work constitutes the largest proteomics dataset for PBMCs min. Sample preparation was performed using the Mass Spec Sample Prep Kit for Cultured Cells (Pierce, Rockford IL). Peptide digests were then analyzed by LC-MS/MS analysis on a Thermo ScientificTM Q ExactiveTM HF mass spectrometer. Database search and oxidation site localization were performed using Thermo ScientificTM Protein Discoverer 2.0. Inferno was then used for further statistical analysis and ProteinCenter was used to extract biological context. Results and Discussion: Access to the complete atlas of gene expressions and posttranslational modifications in PBMCs will permit more sophisticated studies such us the selection of potential biomarkers that could be used for many purposes ranging from diagnostic, prognosis or even help selecting the appropriate therapy for a patient. The bioinformatic analysis of the results yielded the identification of over 8000 proteins. In addition, over 5000 proteins were accurately quantified and over 7000 oxidation events were identified. [Conclusion] Overall, this study not only adds significant value in the mechanistic understanding of redox signalling, but it also creates a valuable protein repository that could lead to the development of new therapeutic strategies. This work constitutes the largest proteomics dataset for PBMCs to date and one of the most comprehensive proteomics warehouse in the clinical proteomics field to date and one of the most comprehensive proteomics warehouse in the clinical proteomics field

Poster.-- 14th Human Proteome Organization World Congress, Vancouver, 27-30 September 2015

Peer reviewed