Table 1.

Pros and cons overview of principal EV isolation methods.

EV Isolation Methods
Method	Pros	Cons
Differential ultracentrifugation (dUC)	Cost-effective [1] Well established and widely employed [134]	Lipoprotein contamination [134] Time consuming [1] Massive EV aggregation and damaging [1]
Density gradient UC	EV subpopulations distinction [1]	Need for operator high training [1] Low final purity [1] Small sample volumes can be processed [1]
Size exclusion chromatography (SEC)	No high-density lipoprotein contamination [111] Serum albumin removal [111] Fast and cheap method [134] EV integrity preservation [1]	Difficulties in EV subpopulations distinction [111] Co-isolation of non-EV particles above the cut-off [1,134]
Immuno-capture	Selective EV isolation from different cell types [111] Easy method, simple instrumentations [111]	Expensive ligands [111] Need for optimization of a mild elution process [111]
Ultrafiltration (UF)	Simple and cheap method [1,111]	Protein contamination [111] Vesicle damaging [1]
Asymmetrial flow field-flow fractionation (AF4)	Gentle fractionation [111] Provides a large range of size-based separation [106] No lipoprotein contamination [111]	Small sample volumes can be processed [106] Co-isolation of particles with the same hydrodynamic size [106]