Coupling size exclusion chromatography to ultracentrifugation improves detection of exosomal proteins from human plasma by LC-MS.
AGO2, Argonaute protein
CSF, cerebrospinal fluid
EVs, extracellular vesicles
Exosomes
FDR, false discovery rate
Human plasma
ILVs, intraluminal vesicles
MVBs, multivesicular bodies
Mass spectrometry
SEC, size exclusion chromatography
Size-exclusion chromatography
UC, ultracentrifugation
Ultracentrifugation
cfNAs, Cell-free nucleic acids
ctDNA, circulating tumor DNA
miRNA, microRNA
Journal
Practical laboratory medicine
ISSN: 2352-5517
Titre abrégé: Pract Lab Med
Pays: Netherlands
ID NLM: 101690848
Informations de publication
Date de publication:
Aug 2021
Aug 2021
Historique:
received:
13
07
2020
revised:
29
04
2021
accepted:
17
06
2021
entrez:
14
7
2021
pubmed:
15
7
2021
medline:
15
7
2021
Statut:
epublish
Résumé
Exosomes are small lipid bilayer vesicles that are defined by their endocytic origin and size range of 30-140 nm. They are constantly produced by different cell types, by both healthy and abnormal cells, and can be isolated from almost all body fluids.Little information exists in isolating exosomes from plasma due to the complexity of its content and the presence of contaminating plasma proteins. We carried-out liquid chromatography-mass spectrometry (LC-MS/MS) analyses of plasma-derived vesicles from 4 healthy donors obtained by 2 coupled methodologies: Ultracentrifugation (UC) coupled with size-exclusion chromatography (SEC) to isolate and subsequently enrich exosomes.We compared the proteins detected by UC alone and UC coupled with SEC. In the coupled UC + SEC methodology we found 52.25% more proteins enriched in exosomes as CD9, Annexins, YWHAZ (14-3-3 family) and others, than by using UC alone. There is also a reduction of 98.8% of contaminating plasma proteins by coupling UC and SEC in comparison to using UC alone. We conclude that exosomes can be successfully isolated from plasma using a very simple combination of standard methods, which could largely improve the proteomics profiling of plasma exosomes.
Identifiants
pubmed: 34258353
doi: 10.1016/j.plabm.2021.e00241
pii: S2352-5517(21)00041-X
pmc: PMC8254000
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e00241Informations de copyright
© 2021 The Authors.
Déclaration de conflit d'intérêts
The authors declare no competing interests.
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