A standardized method for plasma extracellular vesicle isolation and size distribution analysis.

Pubmed ID: 37115777

Pubmed Central ID: PMC10146456

Journal: PloS one

Publication Date: April 28, 2023

MeSH Terms: Reproducibility of Results, Extracellular Vesicles, Chromatography, Gel

Grants: R01 HL102121, R01 HL169390, R21 HL148363, R56 HL161454

Authors: Diehl JN, Ray A, Collins LB, Peterson A, Alexander KC, Boutros JG, Ikonomidis JS, Akerman AW

Cite As: Diehl JN, Ray A, Collins LB, Peterson A, Alexander KC, Boutros JG, Ikonomidis JS, Akerman AW. A standardized method for plasma extracellular vesicle isolation and size distribution analysis. PLoS One 2023 Apr 28;18(4):e0284875. doi: 10.1371/journal.pone.0284875. eCollection 2023.

Studies:

Abstract

The following protocol describes our workflow for isolation and quantification of plasma extracellular vesicles (EVs). It requires limited sample volume so that the scientific value of specimens is maximized. These steps include isolation of vesicles by automated size exclusion chromatography and quantification by tunable resistive pulse sensing. This workflow optimizes reproducibility by minimizing variations in processing, handling, and storage of EVs. EVs have significant diagnostic and therapeutic potential, but clinical application is limited by disparate methods of data collection. This standardized protocol is scalable and ensures efficient recovery of physiologically intact EVs that may be used in a variety of downstream biochemical and functional analyses. Simultaneous measurement quantifies EV concentration and size distribution absolutely. Absolute quantification corrects for variations in EV number and size, offering a novel method of standardization in downstream applications.