Table 5 List of promising modern isolation and separation techniques for MSC-EVs [114]
Technique | Separation system | Advantages | Purity | Sample volume |
|---|---|---|---|---|
Size-exclusion chromatography (SEC) | IZON® qEV column | Removal of co-contaminants including HDLs, albumin Yield better functionality of EVs compared to UC Less compositional and structural alterations comparted to precipitation techniques |  +  +  +  | 100 µl—10 ml |
Sepharose® CL-4B |  +  +  +  | 1 – 10 ml | ||
Filtration-based | Centrifugal filter unit | Defined MWCO ranging from 10 – 100 kDa Simple and easy handling Cost- and time-effective |  +  | Up to 10 ml |
Tangential Flow Filtration (TFF) | Higher concentration of EVs |  +  |  > 10 ml | |
Hydrostatic filtration dialysis (HFD) | No centrifugation step Low EV loss |  +  |  > 10 ml | |
Flow field-flow fractionation | asymmetrical flow field-flow fractionation (AsFlFFF or AF4) | Cross-flow can be modified Optimization between runs possible to enhance separation efficiency More flexible compared to sec Gentle fractionation |  +  +  |  > 10 ml |
Deterministic lateral displacement (DLD) pillar array | Enables separation of exosomes in the size range of 20 to 110 nm |  +  +  |  > 10 ml | |
Charge-based | Anion-exchange chromatography (AIEC) | Shorter isolation time (< 3 h for 1 L of cell culture supernatant) Yield intact evs |  +  +  | Up to 1L |
Electrophoresis and dielectrophoresis (DEP) | Subpopulations separated based on electrophoretic mobilities acquire information on properties of charged and non-charged EVs |  +  +  |  > 10 ml | |
Affinity-based | Magnetic beads | Highly selective and specific Isolate evs originating from different cell types |  +  +  +  | 100 µl–1 ml |