Evaluation of purification methods of EVs

Author: admin     Date: February 20, 2024

One of the challenges that restricts the growth of extracellular vesicle (EV) research field is the lack of a standardized method for EV separation. In this study, three commonly used EV separation methods, ultracentrifugation, precipitation and size exclusion chromatography (combined with ultrafiltration with a microfluidic tangential flow filtration device, Exodisc) was used to comprehensively evaluate the EV yield and sample purity from cell culture medium, human urine and plasma.

The isolated EVs were assessed in terms of their yield, purity and surface protein expression by the Flow NanoAnalyzer. Using these parameter, practical recommendations were made for the choice of isolation and purification methods based on sample type and downstream applications. 

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             Figure 1. Concentrations of particles separated by different methods                   Figure 2.  Purity of EV samples assessed by particle/protein ratio.

 

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                              Figure 3. Composition purity analysis of EVs by DGUC                    Figure 4. Purity of EV samples assessed by single particle phenotyping

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Figure 5. The decision tree on how to select the proper EV separation method based on the input sample.

In this study, the author characterized EVs by the Flow NanoAnalyzer with multiple parameters, evaluated purification methods suitable for different sample types, concluded how to select suitable purification methods based on different sample sources and downstream applications, and provided practical recommendations.

J Extracell Vesicles, 2020, 10(2):e12044.