Table 2 Techniques for EV isolation
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 | Â | Rapidness Purity Efficiency | Complexity of devices Additional equipment Cost |