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Table 2 Techniques for EV isolation

From: Isolation and characterization of extracellular vesicles from Broncho-alveolar lavage fluid: a review and comparison of different methods

Method Processing time Advantages Disadvantages
Differential centrifugation 140–600 min Cost
Isolation from large volumes
Absence of additional chemicals
Equipment (Ultracentrifugation)
Complexity
Efficiency is affected by the type of rotor
Density gradient ultracentrifugation 250 min–2 days Pure preparations
No contamination with viral particles
Absence of additional chemicals
Complexity
Equipment (Ultracentrifugation)
Loss of sample
Size-exclusive chromatography 1 ml/min + column Pure preparations
Preserves vesicle integrity
Prevents EV aggregation
Limitations on sample volume
Specialized equipment and column
Complexity
Commercial kits for polymer precipitation 30–60 min or overnight Simple procedure
No need additional equipment
Cost (especially for diluted samples, such as urine)
Impurities
Precipitation with chemicals (polymers, polyethylene glycol, protamine, sodium acetate) 60–120 min or overnight Cost
Simple procedure
Possibility of processing samples with large volume
Contamination with non-EV proteins
Retention of chemical or polymer
Long duration (sometimes)
Immuno-precipitation (CD9, CD63, CD81 or specific cell type marker) 240 min Purity and high selectivity High selectivity
Cost
Difficulties with detachment of antibodies
Analysis of intact vesicles
Ultrafiltration (nanomembrane or filters with a pore diameter of 0.8–0.1 μm) 130 min Simple procedure
Allowing for concurrent processing of many samples
No limitations on sample volume
Filter plugging (loss sample)
Contamination (proteins)
Microfluidic technologies   Rapidness
Purity
Efficiency
Complexity of devices
Additional equipment
Cost