Volume 5, Issue 3 (2024) – 13 articles
Cover Picture: The miRNA cargo of plasma extracellular vesicles (EVs) is commonly studied for its biomarker potential, but isolation of EVs from human plasma is challenging due to the presence of other non-EV particles. In 2014, we introduced “size-exclusion chromatography” (SEC), a single step isolation method that is commonly used to isolate EVs from plasma and other biofluids.
However, SEC does not completely separate EVs from other miRNA carriers present in plasma, such as cells, lipoproteins, and proteins. Recently, multiple new SEC columns were introduced which differ in resin pore size, column length, etc. Until now, no systematic study has been performed to compare the recovery and purity of plasma EVs using both traditional and new SEC columns.
In this study, we investigated how different SEC columns affect the recovery of plasma EVs (EV yield) and their separation from non-EV miRNA carriers (EV purity). We also investigated how differences in EV recovery and purity affect downstream miRNA analysis, and our results give insight in whether miRNAs are truly associated with plasma EVs. Finally, we investigated column re-use and provide recommendations on the choice of a SEC column.
view this paper However, SEC does not completely separate EVs from other miRNA carriers present in plasma, such as cells, lipoproteins, and proteins. Recently, multiple new SEC columns were introduced which differ in resin pore size, column length, etc. Until now, no systematic study has been performed to compare the recovery and purity of plasma EVs using both traditional and new SEC columns.
In this study, we investigated how different SEC columns affect the recovery of plasma EVs (EV yield) and their separation from non-EV miRNA carriers (EV purity). We also investigated how differences in EV recovery and purity affect downstream miRNA analysis, and our results give insight in whether miRNAs are truly associated with plasma EVs. Finally, we investigated column re-use and provide recommendations on the choice of a SEC column.
