Isolation of Circulating Extracellular Vesicles by High-Performance Size-Exclusion Chromatography.

Circulating extracellular vesicles (EVs) are gaining increased attention as carriers of proteins, nucleic acids, and lipids. Blood contains EVs from different cell sources that constitute an attractive target for biomarker studies. However, there is no consensus on the best approach to isolate EVs from blood. Non-EV proteins and lipoproteins in plasma/serum tend to contaminate isolated EVs and confound functional experiments. Here we describe a single-step, high-performance size-exclusion chromatography procedure for separation of EVs from most lipoproteins and non-EV proteins, and compare it to ultracentrifugation, still the most commonly used method for EV isolation.