Table 1.
Small Extracellular Vesicles (sEVs) Isolation Techniques: Advantages and Disadvantages
| Isolation Technique | Advantages | Disadvantages |
|---|---|---|
| Ultracentrifugation | High recovery rates, simplified operation, no chemical contamination. | Costly instrumentation and time-consuming. Low RNA yields, low purity, and may be mixed with other components. |
| Sucrose density gradient centrifugation | High sEVs purity. | Preparation work is tedious and time-consuming, sEVs levels are low and biological function easily lost. |
| Size-exclusion chromatography | High recovery rates, high purity, less time consuming, high sensitivity, intact structures and biological function preserved. No chemical contamination. It is currently a better method to extract sEVs. | Expensive instrumentation. |
| Ultrafiltration | High efficiency, free from chemical contamination, and low extraction costs. | Low elution efficiency, difficulty in removing the effects of proteins, purity, shape, and charge of the sample affecting the separation. Clogging of filter membranes and loss of biological function. |
| Polymer precipitation | Simple operation, high output, high integrity, no special equipment required, economical. | Cumbersome preparation work, low purity, and susceptibility to chemical contamination. |
| Microfluidics | Easy to operate, high throughput, automated, less sample volume required for highly pure extracted sEVs. | Costly, low yields, and prone to hole plugging. |
| Commercial kits | Easy to operate and short times required. | The kits are expensive, without uniform standards, and are not suitable for the extraction of massive samples. |
| Enzyme-linked immunosorbent assay | High purity, intact isolated sEVs, suitable for the separation of specific sEVs. High specificity. | Antibodies and equipment are costly, low throughput, low isolation efficiency, suitable for cell-free samples only. |
| Hydrostatic filtration dialysis | Low costs, no chemical pollution, sEVs can be isolated from highly diluted solutions, high throughput, high integrity, and biological functions preserved. | Large sample volumes show decreased efficiency |