Updated project metadata. In the present study, we investigated whether sweat-derived extracellular vesicles (EVs) can be used as a source of potential protein biomarkers of human and bacterial origin. We compared protein composition of total sweat, EV-enriched sweat (after ExoEasy) and sweat samples collected with the use of alginate skin patches. We identified 1209 unique human proteins in EV-enriched sweat, of which approximately 20% were present in every individual sample investigated. Besides human proteins, sweat EV samples contained 1594 proteins of bacterial origin. Bacterial protein profiles in EV-enriched sweat were characterized by high interindividual variability, that reflected differences in total sweat composition. Alginate-based sweat patch accumulated about 30% of human proteins from those identified in sweat-derived EVs, but only 5% proteins of bacterial origin. In conclusion, we showed that sweat-derived EVs provide a rich source of potential biomarkers of human and bacterial origin and that commercially available alginate skin patches could be used for collection of human sweat EVs with very little impact of bacterial vesicles.