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

by Alexander, Kyle C. , Boutros, Jacob G. , Peterson, Andrew , Diehl, J. Nathaniel , Ikonomidis, John S. , Ray, Amelia , Akerman, Adam W. , Collins, Lauren B.

in Analysis / Automation / Biology and Life Sciences / Biosynthesis / Chromatography / Chromatography, Gel / Data collection / Ethylenediaminetetraacetic acid / Extracellular matrix / Extracellular Vesicles / Flow cytometry / Health aspects / Lab Protocol / Laboratories / Lipids / Medicine and Health Sciences / Methods / Microscopy / Particle size / People and Places / Physical Sciences / Plasma / Polyethylene glycol / Proteins / Protocol / Reproducibility of Results / Research and Analysis Methods / Science Policy / Size distribution / Size exclusion chromatography / Standardization / Vesicles / Workflow

2023

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

by Alexander, Kyle C. , Boutros, Jacob G. , Peterson, Andrew , Diehl, J. Nathaniel , Ikonomidis, John S. , Ray, Amelia , Akerman, Adam W. , Collins, Lauren B.

2023

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

by Alexander, Kyle C. , Boutros, Jacob G. , Peterson, Andrew , Diehl, J. Nathaniel , Ikonomidis, John S. , Ray, Amelia , Akerman, Adam W. , Collins, Lauren B.

2023

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Journal Article

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

Alexander, Kyle C.,

Boutros, Jacob G.,

Peterson, Andrew,

Diehl, J. Nathaniel,

Ikonomidis, John S.,

Ray, Amelia,

Akerman, Adam W.,

Collins, Lauren B.

2023

Overview

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.

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Publisher

Public Library of Science,Public Library of Science (PLoS)

Subject

Analysis

/ Automation

/ Biology and Life Sciences

/ Biosynthesis

/ Chromatography

/ Chromatography, Gel

/ Data collection