Table 1.

Comparison of EV isolation methods.

Separation method	Principle	Advantages	Disadvantages	Recovery rate	Purity	Time	Sample size	References
Ultracentrifugation	Different sedimentation coefficients for different particles	Suitable for large sample sizes; no other markers are introduced; low cost	High equipment costs; Mechanical damage; long operating time	High	Low	≈4 h	Large	(113, 114)
Density gradient centrifugation	Separation based on particle size, shape, and density	High purity; wide range of applications; Thorough separation of protein aggregates	Cumbersome and time-consuming to operate; contains impurities similar in density to EVs	Low	High	10–18 h	Medium	(115)
Size exclusion chromatography	Chromatographic techniques for separation based on the sizes of sample molecules	Maintains integrity and biological activity; no additional pre-processing required	Potential contamination exists; high equipment costs	High	High	15 min	Small	(116, 117)
Immunoaffinity capture	Isolation of EVs using specific binding between antigen and antibody	High specificity; easy operation	Disruption of EV integrity; higher costs; presence of non-specific binding	Low	High	2–6 h	Small	(118)
Chemical precipitation	Polymers can adhere and precipitate EVs	Wide range of applications; simple operation and high efficiency; less damage to EVs	Low purity; incomplete removal of peak proteins affects proteomic analysis	High	Medium	0.5–12 h	Small	(119)
Microfluidic technology	Combining microfluidics with electrical techniques for EV separation	Maintaining EV integrity; high purity	Not suitable for large samples	Low	High	<2 h	Small	(120, 121)

EV, extracellular vesicle.