Table 1.

Methods of extracellular vesicle isolation.

Isolation method	Working principle	Volume reduction (fold)	EV recovery (%)	Notes
Ultra-centrifugation	Size separation: large EVs collect earlier & at lower g forces	8	10–80	Cost-effective; causes vesicle clumping; pellets soluble factors & ECM along with EVs
Density gradient centrifugation	Separation by density in sucrose/iodixanol gradients	1	5–50	Non-isosmotic sucrose concentrations cause leaching of EV cargoes
Size exclusion chromatography	Column-based size separation	0.2	30–90	Effective separation of plasma proteins; prone to co-isolation of large proteins/aggregates
Ultrafiltration	Pressure-mediated by size/solubility	250	80	EV deformation possible; co-isolation of EV-particle aggregates
Immunocapture	Immobilized antibodies against EV-specific ligands	5	N/A	Cross-reactivity & non-specific binding: recovery of non-EV proteins
Precipitation	PEG polymers exclude water volume	50	90	Inexpensive; concerns re: compatibility with NTA