Summary of commonly used extracellular vesicle (EV) isolation methods
| Technique | Principle | Advantage/disadvantage | Ref |
| Differential ultracentrifugation | EVs are isolated based on their sedimentation velocity in a centrifugal force | Currently most widely used and easy to perform Difficulty achieving full separation of EV subcategories | 148 |
| Size exclusion | Separates EVs based on size | Well characterised kits available Requires extensive postisolation processing | 149 |
| Affinity separation | A molecule with affinity to a surface marker on the EV is suspended in a resin or bead which ‘pulls’ EVs from complex matrices | Can be highly specific for EVs Elution of the EV from the tagged resin requires harsh buffers which can degrade the EV membrane | 150 |
| Precipitation | High molecular weight polymers are complexed with EVs and isolated by either size or density-based methods | Reduces complexity of isolation protocol Commercially available kit—ExoQuick Requires postisolation decomplexation | 53 |
| Density ultracentrifugation | A form of ultracentrifigation (UC) where viscosity gradients are employed to create a cushion which separates molecules into defined layers | Greatly reduces copelleting substances such as cellular lipoproteins Lower EV yields are often reported compared with differential UC | 151 |
| Microfluidic system | Acoustic, electric, optical, magnetic and immunoisolation of EVs from whole material | High purity isolation with qualitative and quantitative potential Low volumes result in lower yield and throughput | 16 |