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Table 1 The techniques for exosome separation

From: Exosomes as a new frontier of cancer liquid biopsy

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 [49, 54]
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 [20, 58, 70]
 Magnetic Separation Antibody-modified magnetic beads Convenient; high efficiency High-cost; marker dependent Separation of exosomes [36, 71,72,73,74]
 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 [62, 76, 77]
Deterministic lateral displacement Continuous accurate and precise separation Low throughout and the requirement of high voltage Efficient isolation of extracellular vesicles [75, 78]
Size-exclusion chromatography High yield; gentle processing Contamination of same-sized vesicles; lack specificity Efficient isolation of extracellular vesicles [50, 52, 79]
 Lipid Mediated-Separation Lipid nanoprobe/TiO2 Minimal damage Contamination of other phospholipid membrane vesicles; lack specificity Efficient isolation of extracellular vesicles [66, 80]
 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 [67, 81]
 Thermophoretic Enrichment Thermophoresis Free from pre-isolation; simple; fast isolation Contamination of same-sized vesicles; lack specificity Efficient isolation of extracellular vesicles [68, 69]