Isolation Defines Identity: Functional Consequences of Extracellular Vesicle Purification Strategies

The biological activity of extracellular vesicles (EVs) is largely defined by their molecular cargo, yet the impact of isolation workflows on EV proteomes and function remains incompletely understood. Here, we compared four isolation strategies for EVs derived from malignant ascites and ES-2 ovarian cancer cell culture supernatants, assessing yield, particle size, protein cargo, and EV-associated enzymatic activity. Proteomic analyses of particle-normalized preparations were performed according to MISEV2023 guidelines, and vesicle-associated protease activity was profiled using a FRET-based assay with inhibitor panels. Principal component and overlap analyses identified a common EV proteome signature for…

• DF
  Deutsche Forschungsgemeinschaft

  Awards: 416910386 GRK 2573, BA1606/4‐2, GRK 2573, DFG-INST 216/793-1 FUGG, KFO325, 416910386, DFG‐INST 216/793‐1 FUGG