Isolation of Extracellular Vesicles (EVs) Using Benchtop Size Exclusion Chromatography (SEC) Columns.

A diverse group of lipid bilayer enclosed nanoparticles, referred to as extracellular vesicles (EVs), are released by all eukaryotic and prokaryotic cells into the extracellular space. The population of EVs being heterogeneous poses a challenge in their efficient separation. Several methods have been employed for EV isolation. However, there is no single conventional method that could recover a high amount of EVs while retaining their purity and functionality. The merging of differential centrifugation with size exclusion chromatography (SEC) is one of the best practices for EV isolation/purification as it recovers a sufficient amount of EVs while retaining their functionality. Here, we describe a method of purification of EVs from bovine follicular fluid samples using benchtop SEC columns. In conclusion, the EV purification method should be chosen based on the downstream applications, as every method poses its own limitations.