CHAPTER 2: Separation and Purification of Extracellular and Bio-fluid Vesicles – State of the Art: Past, Present, and Future of Extracellular Vesicle Separation and Purification
Published:20 Oct 2021
V. Sunkara, H. Woo, and Y. Cho, in Extracellular Vesicles: Applications to Regenerative Medicine, Therapeutics and Diagnostics, ed. W. Chrzanowski, C. T. Lim, and S. Y. Kim, The Royal Society of Chemistry, 2021, pp. 22-44.
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Despite growing interest, there is still no method available for ideal extracellular vesicle (EV) separation with satisfactory yield, purity, and user-friendly operation. Enrichment and purification of EVs are complicated by the fact that EVs are intrinsically heterogeneous in their size, density, and cargo, which makes separation of pure fractions of EVs empirically impossible. Therefore, it is at the discretion of the user to make a strategy for EV separation depending on the sample type, volume, and their downstream applications. Here, basic guidelines are provided to select a method for each type of biofluid, rating each method's performance in terms of total processing time, ease of use, feasible sample volume, yield, and purity of the concentrated EVs. The emerging technologies to achieve pure EVs and to unveil their heterogeneity are outlined together with the remaining challenges that need to be overcome.