The efficiency of anticancer therapy depends heavily on the molecular features of the tumor. Since tumor-derived extracellular vesicles, including exosomes, contain proteins that are characteristic for producer cells and could be detected in biological fluids, they are of great interest for screening of predictive biomarkers. We have performed label-free mass spectrometric profiling of extracellular vesicles originated from human colon cancer cell lines Caco-2, HT-26 and HCT-116, and from lung cancer cell lines NCI-H23 and A549.