Skip to main content

Table 2 Summary of common EV isolation methods

From: Basics and applications of tumor-derived extracellular vesicles

Method Description Example Yield Purity/quality Equipment/Cost Approximate processing time Commercial products Reference
Ultracentrifugation-based methods Size- or density-dependent isolation 1. Differential centrifugation/Ultracentrifugation 2. Sucrose gradient ultracentrifugation High Protein aggregates and particulates contaminations High equipment cost 2.5 – 48 h No commercial available [75, 77,78,79,80]
Size-based methods Size- or molecular weight-dependent isolation 1. Ultrafiltration 2. Size exclusive chromatography (SEC) 3. Asymmetrical-flow field-flow fractionation (AF4) Low (AF4) High purity (AF4, SEC); exosomes may be deformed (ultrafiltration) Low/moderate equipment cost (ultrafiltration and SEC ); high equipment cost (AF4) 1 - 1.5 h (Ultrafiltration); 0.5 - 1 h (SEC); 1h (AF4) Available, .e.g.. EVSecond column (GL Sciences) or qEV column (Izon Science) [82,83,84, 98,99,100]
Immunoaffinity-based methods Antibody-antigen interaction-dependent isolation 1. Immunocapture 2. Immunoadsorption Low High purity High reagent cost 4 - 5 h Available, e.g. MagaCapture™ Exosome Isolation Kit (Wako) or Exosome-Human CD9 Isolation Reagent (ThermoFisher) [85,86,87]
Precipitation-based methods Solubility- or dispersibility-dependent isolation Polyethylene glycol Dependence Contaminated precipitates Low equipment cost 1 h Available, e.g. Total Exosome Isolation Kit (Invitrogen) or ExoQuick™ Exosome Precipitation (System Biosciences) [81, 88, 89]
Microfluidic-based methods Size-, density-, or antibody-antigen interaction-dependent isolation Microfluidic device with nanoporous membrane, immuno-chip, or porous silicon nanowires-on-micropillar structure Dependence High purity (porous nanowires-on-micropillar structure); EVs may be damaged (nanoporous membrane-based filtration) Low/moderate equipment cost 2 h (porous membrane-based filtration); 1.5 h (immuno-chip) No commercial available [91,92,93,94,95]
Flow cytometry-based methods Antibody-antigen interaction-dependent isolation 1. Fluorescent-labeled antibody-based isolation 2. Immuno-magnetic-based isolation Low High purity High equipment and reagent cost 12 h Avaialbe, e.g. Exosome Flow Cytometry Kit (Wako) [96, 97]