Table 1.
Overview of the main methods to isolate urinary vesicles.
| Reference | Short method description | Advantages | Disadvantages | |
|---|---|---|---|---|
| Barutta et al. (69) | Differential centrifugation | Vesicles enrichment as a pellet | No fixed conditions for: 1. number of centrifugations; 2. relative centrifugation force; 3. time; 4. rotor type; 5. sample volume; 6. temperature during centrifugation; and 7. presence/absence of protease inhibitors | Not applicable for large volume of samples and not suitable for large cohort of patients. Expensive equipment like ultracentrifuge and HPLC are necessary to implement UEVs enrichment |
| Gildea et al. (47) | Differential centrifugation | |||
| Kalani et al. (72) | Differential centrifugation | |||
| Lv et al. (65) | Differential centrifugation | |||
| Musante et al. (98) | Differential centrifugation + CHAPS treatment | Protein activity prevention | Labor intensive | |
| Gonzales et al. (54) | Differential centrifugation + DTT treatment | Increase of the exosomal yield in the ultracentrifugation pellet from THP interference | Not suitable for protein activity assessment designated samples. Incomplete removal of THP from the ultracentrifugation pellet | |
| Fernandez-Llama et al. (95) | Differential centrifugation + DTT treatment | |||
| Wang et al. (55) | Differential centrifugation + DTT treatment | |||
| Cheng et al. (63) | Differential centrifugation + DTT treatment | |||
| Rood et al. (91) | Differential centrifugation + SEC | Vesicle separation according to size | Labor intensive | |
| Rood et al. (91) | Nano-membrane filtration | Concentration and removal of soluble proteins | Differences in removal of bigger (>0.22 μm or more) vesicles without assessment of their importance for biomarkers screening. Adsorption of vesicles on the surface of ultrafiltration membrane | |
| Merchant et al. (89) | Differential centrifugation + microfiltration | |||
| Miranda et al. (62) | Differential centrifugation + nanofiltration | |||
| Principe et al. (57) | Differential centrifugation + ultrafiltration | |||
| Prunotto et al. (59) | Differential centrifugation + ultrafiltration | |||
| Hogan et al. (52) | Differential centrifugation + sucrose gradient | Vesicle separation according to density | Extremely labor intensive | |
| Raimondo et al. (116) | Differential centrifugation + sucrose gradient | |||
| Ramirez-Alvarado et al. (90) | Ultrafiltration + differential centrifugation | Vesicle separation according to density | Extremely labor intensive | |
| Zubiri et al. (77) | Exoquick | No need of ultracentrifugation step. Suitable for extraction of RNA and DNA | Precipitation solution is expensive especially when applied for large volumes of urine and number of samples. For proteomic analysis, extra steps are necessary to remove the interference from precipitating agent | |
| Alvarez (64) | Exoquick | |||
| Musante et al. (unpublished) | Hydrostatic dialysis | Inexpensive, large volume of urine | ||