Modern isolation and separation techniques for extracellular vesicles

• Published in: Journal of Chromatography A Vol. 1636; p. 461773
• Main Authors: Liangsupree, Thanaporn, Multia, Evgen, Riekkola, Marja-Liisa
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 11.01.2021
• Subjects: Affinity-based; automation; Charge-based; chromatography; Chromatography, Affinity; Chromatography, Gel; Chromatography, Ion Exchange; Electrophoresis; Extracellular vesicles; Extracellular Vesicles - chemistry; fractionation; Fractionation, Field Flow; Humans; Isolation; Separation; Size-based; therapeutics; Ultracentrifugation; Extracellular vesicles; Isolation; Separation; Size-based; Charge-based; Affinity-based
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2020.461773

•Modern techniques for extracellular vesicle isolation and separation are reviewed.•Emphasis is given to modern size-, charged-, and affinity-based approaches.•A trend is toward the combination of multiple techniques for EV isolation.•Studies of EV subpopulations require combinations of techniques.•Automated on-line technique allows quick and reliable EV subpopulation studies. Extracellular vesicles (EVs) are heterogenous membrane-bound vesicles released from various origins. EVs play a crucial role in cellular communication and mediate several physiological and pathological processes, highlighting their potential therapeutic and diagnostic applications. Due to the rapid increase in interests and needs to elucidate EV properties and functions, numerous isolation and separation approaches for EVs have been developed to overcome limitations of conventional techniques, such as ultracentrifugation. This review focuses on recently emerging and modern EV isolation and separation techniques, including size-, charge-, and affinity-based techniques while excluding ultracentrifugation and precipitation-based techniques due to their multiple limitations. The advantages and drawbacks of each technique are discussed together with insights into their applications. Emerging approaches all share similar features in terms of being time-effective, easy-to-operate, and capable of providing EVs with suitable and desirable purity and integrity for applications of interest. Combination and hyphenation of techniques have been used for EV isolation and separation to yield EVs with the best quality. The most recent development using an automated on-line system including selective affinity-based trapping unit and asymmetrical flow field-flow fractionation allows reliable isolation and fractionation of EV subpopulations from human plasma.