Table 1 Isolation methods of sEVs
From: Current aspects of small extracellular vesicles in pain process and relief
Methods | Process | Advantages | Disadvantages | Ref. | |
|---|---|---|---|---|---|
Centrifugation | Differential ultracentrifugation | 400×g, 2000×g, 10,000×g centrifugation, and then ultracentrifugation of more than 100,000×g to obtain sEVs | Simple process, moderate production | Required expensive device, impurity, time-consuming | |
Density gradient centrifugation | DUC combines with separation media | High purity | Complicated process, low yield | [37] | |
Vesicle-size separation | Size-exclusion chromatography | Vesicles of different sizes were separated by different velocities in the filling process of porous polymer gels | High purity | Suitable for small-volume liquid, low yield | [34] |
Ultrafiltration | Appropriate pore size is used to eliminate larger cell debris and cells, thereby obtaining sEVs on the membrane | High yield, simple process, rapid | Low purity, larger vesicles | [38] | |
Immunoaffinity isolation | Immunomagnetic separation | The antibodies on beads combine with antigens on vesicles to capture sEVs | Specific vesicles, high purity | Expensive, low yield | [39] |
Device | Microfluidic | precise control of samples and vesicle collection through miniaturized devices and microvalves, facilitating dynamic manipulation and isolation of sEVs | Precise control, stable yield, automation process | Expensive microfluidic device, low-volume sample | [40] |
Others | Commercial kits | According to the commercial protocol | Simple process | Expensive, unstable quality | [41] |