In:
PLOS ONE, Public Library of Science (PLoS), Vol. 18, No. 5 ( 2023-5-10), p. e0285440-
Abstract:
Extracellular vesicles (EVs) isolated from plasma are increasingly recognized as promising circulating biomarkers for disease discovery and progression, as well as for therapeutic drug delivery. The scientific community underlined the necessity of standard operative procedures for the isolation and storage of the EVs to ensure robust results. The understanding of the impact of the pre-analytical variables is still limited and some considerations about plasma anticoagulants and isolation methods are necessary. Therefore, we performed a comparison study between EVs isolated by ultracentrifugation and by affinity substrate separation from plasma EDTA and sodium citrate. The EVs were characterized by Nano Tracking Analysis, Western Blot, cytofluorimetric analysis of surface markers, and lipidomic analysis. While anticoagulants did not significantly alter any of the analyzed parameters, the isolation methods influenced EVs size, purity, surface markers expression and lipidomic profile. Compared to ultracentrifugation, affinity substrate separation yielded bigger particles highly enriched in tetraspanins (CD9, CD63, CD81), fatty acids and glycerolipids, with a predominant LDL- and vLDL-like contamination. Herein, we highlighted that the isolation method should be carefully evaluated prior to study design and the need of standardized operative procedures for EVs isolation and application to biomarkers discovery.
Type of Medium:
Online Resource
ISSN:
1932-6203
DOI:
10.1371/journal.pone.0285440
DOI:
10.1371/journal.pone.0285440.g001
DOI:
10.1371/journal.pone.0285440.g002
DOI:
10.1371/journal.pone.0285440.g003
DOI:
10.1371/journal.pone.0285440.g004
DOI:
10.1371/journal.pone.0285440.g005
DOI:
10.1371/journal.pone.0285440.t001
DOI:
10.1371/journal.pone.0285440.s001
DOI:
10.1371/journal.pone.0285440.s002
DOI:
10.1371/journal.pone.0285440.s003
DOI:
10.1371/journal.pone.0285440.s004
DOI:
10.1371/journal.pone.0285440.s005
DOI:
10.1371/journal.pone.0285440.s006
DOI:
10.1371/journal.pone.0285440.s007
DOI:
10.1371/journal.pone.0285440.s008
DOI:
10.1371/journal.pone.0285440.s009
Language:
English
Publisher:
Public Library of Science (PLoS)
Publication Date:
2023
detail.hit.zdb_id:
2267670-3
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