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They carry proteins and RNAs, both miRNAs and mRNAs, and have been shown to transfer their cargo to recipient cells 3, 8, 9. Exosomes are 70–150 nm in size and originate from the endocytic pathway 5 whereas MVs are generally larger, 100–1000 nm in diameter and bud directly from the plasma membrane 6, 7. In this article, the term EVs will refer to exosomes and MVs only. Hence, the BE-SEC based EV purification method represents an important methodological advance likely to facilitate robust and reproducible studies of EV biology and therapeutic application.Įxtracellular vesicles (EVs) are nanosized cell-derived vesicles 1, 2, 3 delimited by a lipid bilayer and typically divided into three subgroups, according to their biogenesis pathways exosomes, microvesicles (MVs) and apoptotic bodies 4. Furthermore, uptake of eGFP labelled EVs in recipient cells was comparable between BE-SEC and UC samples. This technique is reproducible and scalable, and surface marker analysis by bead-based flow cytometry revealed highly similar expression signatures compared with UC-purified samples. Here we show that commercially available bind-elute size exclusion chromatography (BE-SEC) columns purify EVs with high yield (recovery ~ 80%) in a time-efficient manner compared to current methodologies.

EVs have traditionally been purified by ultracentrifugation (UC), however UC has limitations, including resulting in, operator-dependant yields, EV aggregation and altered EV morphology, and moreover is time consuming. Extracellular vesicles (EVs) play a pivotal role in cell-to-cell communication and have been shown to take part in several physiological and pathological processes.