Rapid Nickel-based Isolation of Extracellular Vesicles from Different Biological Fluids.
Biomarkers
Extracellular vesicles
Liquid biopsies
Nickel
OMVs
Journal
Bio-protocol
ISSN: 2331-8325
Titre abrégé: Bio Protoc
Pays: United States
ID NLM: 101635102
Informations de publication
Date de publication:
05 Feb 2020
05 Feb 2020
Historique:
received:
25
09
2019
revised:
19
12
2019
accepted:
20
12
2019
entrez:
3
3
2021
pubmed:
4
3
2021
medline:
4
3
2021
Statut:
epublish
Résumé
Extracellular vesicles (EVs) are membranous structures that cells massively release in extracellular fluids. EVs are cargo of cellular components such as lipids, proteins, and nucleic acids that can work as a formidable source in liquid biopsy studies searching for disease biomarkers. We recently demonstrated that nickel-based isolation (NBI) is a valuable method for fast, efficient, and easy recovery of heterogeneous EVs from biological fluids. NBI exploits nickel cations to capture negatively charged vesicles. Then, a mix of balanced chelating agents elutes EVs while preserving their integrity and stability in solution. Here, we describe steps and quality controls to functionalize a matrix of agarose beads, obtain an efficient elution of EVs, and extract nucleic acids carried by them. We demonstrate the versatility of NBI method in isolating EVs from media of primary mouse astrocytes, from human blood, urine, and saliva processed in parallel, as well as outer membrane vesicles (OMVs) from cultured Gram-negative bacteria.
Identifiants
pubmed: 33654737
doi: 10.21769/BioProtoc.3512
pii: e3512
pmc: PMC7842538
doi:
Types de publication
Journal Article
Langues
eng
Pagination
e3512Informations de copyright
Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.
Déclaration de conflit d'intérêts
Competing interestsMN, AP, AQ, and VGD declare a potential conflict of interest for a patent application regarding the NBI method (P019950WO).
Références
J Cell Sci. 2019 Jul 1;132(13):
pubmed: 31263077
Clin Chim Acta. 2019 Jun;493:14-19
pubmed: 30797770
Am J Physiol Renal Physiol. 2014 Jun 1;306(11):F1251-9
pubmed: 24694589
Curr Protoc Cell Biol. 2006 Apr;Chapter 3:Unit 3.22
pubmed: 18228490
J Cell Biol. 2013 Feb 18;200(4):373-83
pubmed: 23420871
Sci Rep. 2018 Apr 10;8(1):5730
pubmed: 29636530
Nat Rev Mol Cell Biol. 2018 Apr;19(4):213-228
pubmed: 29339798
Exp Mol Med. 2019 Mar 15;51(3):1-10
pubmed: 30872566
Anal Bioanal Chem. 2019 Aug;411(21):5351-5361
pubmed: 31267193
J Transl Med. 2011 Jan 14;9:9
pubmed: 21235781
Adv Clin Chem. 2016;76:37-53
pubmed: 27645815
Mol Cell Proteomics. 2017 Jul;16(7):1348-1364
pubmed: 28483926
J Extracell Vesicles. 2019 Apr 04;8(1):1596669
pubmed: 31007875
Cell. 2019 Apr 4;177(2):428-445.e18
pubmed: 30951670
J Biol Chem. 2013 May 31;288(22):15699-711
pubmed: 23592792
J Proteomics. 2019 Jul 30;204:103403
pubmed: 31170500
EBioMedicine. 2019 May;43:114-126
pubmed: 31047861
Sci Rep. 2016 Aug 09;6:31175
pubmed: 27503761
Proc Natl Acad Sci U S A. 2016 Feb 23;113(8):E968-77
pubmed: 26858453
J Extracell Vesicles. 2018 Nov 23;7(1):1535750
pubmed: 30637094
Biochim Biophys Acta. 2014 Dec;1846(2):539-46
pubmed: 25444714
Biomed Res Int. 2015;2015:152926
pubmed: 26583089
Sci Rep. 2015 Nov 30;5:17319
pubmed: 26616523
Anal Chem. 2018 Nov 6;90(21):12783-12790
pubmed: 30346136
Cold Spring Harb Protoc. 2010 Jun;2010(6):pdb.prot5438
pubmed: 20516176