A comparison of methods for the isolation and separation of extracellular vesicles from protein and lipid particles in human serum

Title: A comparison of methods for the isolation and separation of extracellular vesicles from protein and lipid particles in human serum
Authors: Brennan, KieranMartin, K.FitzGerald, S. P.Wu, Y.Blanco-Fernandez, AlfonsoMc Gee, Margaret M.et al.
Permanent link: http://hdl.handle.net/10197/11783
Date: 23-Jan-2020
Online since: 2020-12-04T13:12:56Z
Abstract: Extracellular vesicles (EVs) are nano-sized vesicles containing nucleic acid and protein cargo that are released from a multitude of cell types and have gained significant interest as potential diagnostic biomarkers. Human serum is a rich source of readily accessible EVs; however, the separation of EVs from serum proteins and non-EV lipid particles represents a considerable challenge. In this study, we compared the most commonly used isolation techniques, either alone or in combination, for the isolation of EVs from 200 µl of human serum and their separation from non-EV protein and lipid particles present in serum. The size and yield of particles isolated by each method was determined by nanoparticle tracking analysis, with the variation in particle size distribution being used to determine the relative impact of lipoproteins and protein aggregates on the isolated EV population. Purification of EVs from soluble protein was determined by calculating the ratio of EV particle count to protein concentration. Finally, lipoprotein particles co-isolated with EVs was determined by Western blot analysis of lipoprotein markers APOB and APOE. Overall, this study reveals that the choice of EV isolation procedure significantly impacts EV yield from human serum, together with the presence of lipoprotein and protein contaminants.
Funding Details: Irish Research Council
University College Dublin
Funding Details: The Wellcome Trust (WT)
Type of material: Journal Article
Publisher: Springer
Journal: Scientific Reports
Volume: 10
Copyright (published version): 2020 the Authors
Keywords: Extracellular signalling moleculesMultivesicular bodies
DOI: 10.1038/s41598-020-57497-7
Language: en
Status of Item: Peer reviewed
ISSN: 2045-2322
This item is made available under a Creative Commons License: https://creativecommons.org/licenses/by-nc-nd/3.0/ie/
Appears in Collections:Conway Institute Research Collection
Biomolecular and Biomedical Science Research Collection

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