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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]