TABLE 2.

Typical strategies and examples of applications to prepare PDEVs.

Strategies	Isolation condition	Advantage	Disadvantage	References
Ultracentrifugation	A range of centrifugation with different speeds and gradient ultracentrifugation. Considered as the gold standard in EVs extraction because of its capacity to extract EVs in a relatively high-purity fashion	Fast procedure; no limitations on sample volume; pure preparations; low cost	Vesicles trapping; loss of PDEVs; the possibility of clogging	Zhang et al. (2021)
Precipitation	Combination of centrifugation and clustering agents like PEG 6000 to trap EVs	The simplicity of the procedure, low cost, preservation of PDEVs integrity	Retention and contamination of the polymer	Jia et al. (2022)
Immunoaffinity	Using the interaction between the antigens on the surface of EVs and matching antibodies	Purity, high selectivity; convenient operation, low cost	High reagent cost; low yields; difficult to analyze complete vesicles	Sundaram et al. (2020)
Size-based isolation (SEC)	Separating similar-sized particles with ultrafiltration, size-exclusion chromatography, and flow field-flow fractionation, which typically resulted only in an EVs-enriched sample	Purity; reproducibility; preserves vesicle integrity; scalability; prevents PDEVs aggregation	Complexity; limitations on sample volume; specialized equipment; a small number of samples; high cost	Sidhom et al. (2020b)
Commercial isolation kit	EVs isolation kit	Suitable for small samples, simple steps, fast procedure	Impurity; low production; expensive reagents	Muraoka et al. (2020)