Table 1 Summary of extracellular vesicles isolation techniques
From: Application of engineered extracellular vesicles for targeted tumor therapy
Technique | Principle | Advantages | Disadvantages | Isolation capacity | Purity | Time required | References |
|---|---|---|---|---|---|---|---|
Ultracentrifugation (UC) | Particles of different sizes and densities are separated using different speeds | •Gold standard for isolation •Suitable for large-volume samples | • Dependence on equipment • Low purity • Protein aggregation • EVs can be destroyed by high-speed centrifugation | Low | Low | Medium | |
Density gradient centrifugation | Using differences in particle density to maintain different particles in the proper density medium and perform centrifugation | • The purity of EVs is higher than that obtained through UC • Applicable to the isolation of EV subpopulations | • Dependence on equipment • Time-consuming • Risk of EV destruction | Medium | Medium | High | |
Ultrafiltration | Utilizes differences in EV particle size and molecular weight | • Easy to operate • Direct extraction of RNA • Low equipment cost • Good portability | • Insufficient specificity • May contain impurities • Moderate purity | Medium | Medium | High | |
Size exclusion chromatography (SEC) | Utilizes differences in EV particle size and molecular weight | • Preservation of EV structural integrity • Easy to operate • Higher purity than that obtained through UC | • Time-consuming • Possibility of pore blockage • May contain impurities • High device costs | Low | Medium | High | |
Polymer precipitation | Precipitation of EVs through reduced polymer solubility | • Equipment-independent method • High efficiency •Can be used for large samples • Easy to operate | • Easy of contamination • Insufficient specificity • Protein aggregation | Low | Medium | High | |
Immunoaffinity chromatography | Specific antibodies bind to proteins on the surface membrane of EVs | • Higher purity than that obtained through UC • No chemical contamination • Suitable for isolating EVs with identical membrane proteins | • High cost • Can influence EV activity • EV markers must be optimized • Unsuitable for isolation from large samples | Medium | Medium | High | |
Microfluidic technology | Microscale technique using equipment based on physicochemical differences in EVs | • Simplicity and efficiency • Ease of automation and integration • High sensitivity and higher purity compared with that obtained through UC | • Requirement for complicated equipment • Lack of uniform standards | High | Medium | High |