Techniques | Methods | Advantages | Disadvantages | Prominent examples | Ref. |
---|---|---|---|---|---|
Conventional techniques | |||||
 Ultracentrifugation-based Separation | Differential ultracentrifugation | High purity; established protocol; | Lengthy process; large sample volume; requires ultracentrifuge | Separation of EVs from reticulocyte culture medium | [44] |
Gradient density ultracentrifugation | High purity; | Lengthy process; large sample volume; requires ultracentrifuge | Sucrose gradient-purified prostasomes | [46] | |
 Size-based Separation | Ultracentrifugation with ultrafiltration | High purity; high yield | Contamination of same-sized vesicles; lack specificity; difficulty in scaling | Separation of urinary exosomes | [49] |
size-exclusion chromatography | High yield; gentle processing | Contamination of same-sized vesicles; lack specificity; difficulty in scaling | Isolation of EVs from platelet-free supernatant of platelet concentrates | [50] | |
 Precipitation | Polyethylene glycol precipitation | Simple; fast isolation | Lack specificity; much contamination; difficulty in scaling | Isolation of exosomes from plasma, cell culture supernatant | |
Commercial kits | Simple; fast isolation | Lack specificity; much contamination; high price | Isolation of exosomes from serum and/or plasma | [56] | |
Novel techniques | |||||
 Immunoaffinity Enrichment | Antibody-conjugated platform | Simple; specificity | High-cost; marker dependent | Enrichment of exosomes from clinical samples | |
 Magnetic Separation | Antibody-modified magnetic beads | Convenient; high efficiency | High-cost; marker dependent | Separation of exosomes | |
 Physical Feature-based separation | Nanoscale lateral displacement | Reduced membrane blockage; gentle processing | Contamination of same-sized vesicles; lack specificity | On-chip sorting and quantification of exosomes | [75] |
Membrane filter | Gentle processing | Contamination of same-sized vesicles; lack specificity | On-chip isolation of intact extracellular vesicles | ||
Deterministic lateral displacement | Continuous accurate and precise separation | Low throughout and the requirement of high voltage | Efficient isolation of extracellular vesicles | ||
Size-exclusion chromatography | High yield; gentle processing | Contamination of same-sized vesicles; lack specificity | Efficient isolation of extracellular vesicles | ||
 Lipid Mediated-Separation | Lipid nanoprobe/TiO2 | Minimal damage | Contamination of other phospholipid membrane vesicles; lack specificity | Efficient isolation of extracellular vesicles | |
 Acoustic-based microfluidics | Aacoustic radiation force (ARF) and dielectrophoretic (DEP) | Contact-free; high-throughput; continuous separation; wide range of particles | Design and fabrication finer gradations; finer-grade separation of subpopulations | Active sorting of extracellular vesicles | |
 Thermophoretic Enrichment | Thermophoresis | Free from pre-isolation; simple; fast isolation | Contamination of same-sized vesicles; lack specificity | Efficient isolation of extracellular vesicles |