Table 3.

Major isolation technology of EVs/exo.

Isolation technique	Advantage	Disadvantage
Differential ultracentrifugation	The gold standard isolation approach suitable for large-volume specimens, especially cell culture supernatant, urine, etc. low cost	lengthy timescales; the requirement of a large number of cells or biological fluids; non-vesicular macromolecule contamination
Density-gradient centrifugation	High purity of EVs; preservation of EV activity	Time and labor consuming; expensive equipment requirements; large amount of pre-work and complicated steps; low EV yield
Ultrafiltration	Short operation time; low equipment cost; suitable for large-volume specimens	Labor consuming; moderate purity; possible loss due to clogging membrane; potential physical changes induced by shear stress; low EV yield
Immunoaffinity capture	High specificity and purity; easy operation; no potential mechanical damage	High-cost; low efficiency; environment-susceptive ligands activity susceptible to processing environment; low EV processing volume and yields
Size-exclusion chromatography	High purity; easy operation; good reproducibility; preservation of the native state of EVs; suitable for most of downstream analysis	Time consuming; relatively high preparation cost
Polymer Precipitation	Short operation time; preservation of the native state of EVs; high yield; easy accessibility	Contaminations of non-EVs; affection on downstream analysis and quantification of EV samples
Microfluidic techniques	Low sample consumption; fast processing time; high sensitivity; suitable for quantitative detection of scarce samples	Low sample capacity; nonspecific binding