Diego Baranda Martinez Abascal - Secondment from VHIR to NKUA

Diego Baranda from partner VHIR completed his secondment at the National and Kapodistrian University of Athens, as part of the EXPAND-EV project and shared his insights and impressions on this experience.

"I had such a productive stay during October and November at the proteomics team leaded by Makis Zoidakis.

Using my own saliva as a training sample, I learnt the standard methodology for processing of samples for analysis by Mass Spectrometry (MS), consisting on concentration of the sample, complete lysis and tryptic digestion and purification of proteins. I also received complete training in bioinformatic analysis of raw proteomic data obtained from the MS. I learnt how to calculate replicates variability coefficients in order to determine reliability of results as well as the options for proteomes thorough comparison and results display in graphs.

After finishing my training, I screened different methodologies for Extracellular Vesicle (EV) processing along with my colleague Iwona Borošak. The method that showed best simplicity-results quality ratio was selected for further experiments, where small details were polished. At the end we got an optimized Sample Operating Procedure for proteomic analysis of EVs, that will serve as the reference methodology that the other consortium members will use to prepare EVs for proteomic analysis at NKUA.

Finally, I also could take advantage of my time there to analyze a variety of samples I brought from VHIR. I obtained some data for the Expand-EV project with the proteome of both bulk and Cancer Stem Cells subpopulation from two different breast cancer cell lines. Furthermore, I could perform some additional analysis of different EV models produced in the mark of VHIR projects that bear recombinant proteins for potential therapeutic use."