Size-exclusion chromatography for the characterization of urinary extracellular vesicles.
Centrifugation
EVs
Extracellular vesicles
Size-exclusion chromatography
Urine
Journal
Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
ISSN: 1873-376X
Titre abrégé: J Chromatogr B Analyt Technol Biomed Life Sci
Pays: Netherlands
ID NLM: 101139554
Informations de publication
Date de publication:
01 Aug 2023
01 Aug 2023
Historique:
received:
01
04
2023
revised:
07
07
2023
accepted:
12
07
2023
pmc-release:
01
08
2024
medline:
12
9
2023
pubmed:
23
7
2023
entrez:
22
7
2023
Statut:
ppublish
Résumé
In recent years, extracellular vesicles (EVs) have gained attention for their potential as biomarkers for the early diagnosis and treatment of various diseases. Traditionally, EV isolation has relied exclusively on ultracentrifugation. However, alternative enrichment methods such as size-exclusion chromatography (SEC) and polyethylene glycol-based precipitation have been introduced. This study utilized SEC as a characterization tool to assess the efficiency of EV isolation. Urinary EVs isolated from human urine using centrifugation (40,000 × g) were analyzed using an SEC column with a pore size of 1000 Å, an inner diameter of 7.8 mm, and a length of 300 mm. The EVs were detected sequentially using UV (280 nm) and fluorescence (λex/em = 550 nm/565 nm); the EVs were observed at approximately 6 min, while the proteins were observed at approximately 12 min. The repeated centrifugation enrichment steps resulted in an increase in EV peaks and a decrease in protein peaks. SEC analysis of the enriched EV samples confirmed that a four-cycle repetition of centrifugation is necessary for successful EV enrichment and removal of non-EV proteins from 40 mL of human urine.
Identifiants
pubmed: 37480686
pii: S1570-0232(23)00238-6
doi: 10.1016/j.jchromb.2023.123828
pmc: PMC10530618
mid: NIHMS1926712
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
123828Subventions
Organisme : NCI NIH HHS
ID : R01 CA258240
Pays : United States
Informations de copyright
Copyright © 2023 Elsevier B.V. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Références
J Extracell Vesicles. 2021 May;10(7):e12093
pubmed: 34035881
J Extracell Vesicles. 2019 Apr 01;8(1):1596016
pubmed: 30988894
Signal Transduct Target Ther. 2020 Aug 3;5(1):144
pubmed: 32747657
Sci Rep. 2016 Apr 12;6:23978
pubmed: 27068479
J Proteome Res. 2018 Oct 5;17(10):3599-3605
pubmed: 30192545
Anal Chem. 2022 Oct 11;94(40):13676-13681
pubmed: 36166570
Anal Chem. 2018 Jul 17;90(14):8314-8319
pubmed: 29920066
Proteomics. 2023 May;23(9):e2200364
pubmed: 36624553
Pharmacol Ther. 2017 Jun;174:63-78
pubmed: 28202367
Front Pharmacol. 2020 Jan 31;10:1655
pubmed: 32082158
Stem Cells Int. 2022 Sep 6;2022:3577015
pubmed: 36110890
PLoS One. 2012;7(3):e33037
pubmed: 22448232
Proteomics. 2013 Nov;13(22):3354-64
pubmed: 24115447
Biochem Soc Trans. 2013 Feb 1;41(1):241-4
pubmed: 23356290
J Proteome Res. 2020 Jun 5;19(6):2217-2225
pubmed: 32248692
Pharmaceutics. 2023 Jan 21;15(2):
pubmed: 36839687
Biochim Biophys Acta Rev Cancer. 2019 Apr;1871(2):455-468
pubmed: 31047959
Lab Med. 2022 Jan 6;53(1):6-11
pubmed: 34350959
Anal Bioanal Chem. 2019 Jul;411(18):4141-4149
pubmed: 30762101
Cancers (Basel). 2021 Mar 26;13(7):
pubmed: 33810357
Proc Natl Acad Sci U S A. 2014 May 20;111(20):7208-11
pubmed: 24799690
Front Cell Dev Biol. 2018 Feb 20;6:18
pubmed: 29515996
J Chromatogr A. 2021 Jan 11;1636:461773
pubmed: 33316564
Int Immunol. 2005 Jul;17(7):879-87
pubmed: 15908444
Proteomics. 2013 May;13(10-11):1572-80
pubmed: 23129434
Proteomics. 2013 May;13(10-11):1667-1671
pubmed: 23533145
Anal Chim Acta. 2021 Aug 29;1175:338633
pubmed: 34330441
Kidney Int Rep. 2017 Dec 01;3(3):555-572
pubmed: 29854963
Methods Mol Biol. 2017;1660:191-199
pubmed: 28828657
Appl Biochem Biotechnol. 2016 Feb;178(3):544-57
pubmed: 26490380
J Control Release. 2021 Jan 10;329:894-906
pubmed: 33058934
Nature. 2015 Nov 19;527(7578):329-35
pubmed: 26524530
Mol Cell Biochem. 2019 Sep;459(1-2):1-6
pubmed: 31073888
Front Immunol. 2023 Jan 17;13:1093607
pubmed: 36733388
Anal Chem. 2019 Oct 15;91(20):13297-13305
pubmed: 31549806
Langmuir. 2018 Aug 14;34(32):9387-9393
pubmed: 29542322
Electrophoresis. 2015 Sep;36(17):2017-26
pubmed: 26010067
Rapid Commun Mass Spectrom. 2023 Jan 30;37(2):e9427
pubmed: 36321680
J Transl Med. 2018 Jan 9;16(1):1
pubmed: 29316942
Cancer Res. 2017 Dec 1;77(23):6480-6488
pubmed: 29162616
Crit Rev Oncol Hematol. 2022 Apr;172:103628
pubmed: 35189326
J Proteome Res. 2009 Mar;8(3):1304-14
pubmed: 19199708
Mass Spectrom Rev. 2023 Mar;42(2):844-872
pubmed: 34747512
J Circ Biomark. 2015 Jul 10;4:6
pubmed: 28936242
J Proteomics. 2013 Apr 26;82:193-229
pubmed: 23376485
Pharmazie. 2021 Feb 25;76(2):61-67
pubmed: 33714281
ScientificWorldJournal. 2015;2015:657086
pubmed: 25695100
Infect Agent Cancer. 2023 Jan 19;18(1):3
pubmed: 36658631
Proteomics. 2011 Jun;11(12):2459-75
pubmed: 21595033
Mass Spectrom Rev. 2011 Nov-Dec;30(6):1185-202
pubmed: 21544848