Molecular evaluation of five different isolation methods for extracellular vesicles reveals different clinical applicability and subcellular origin.

Autor: Veerman RE; Department of Clinical Immunology and Transfusion Medicine and Division of Immunology and Allergy, Department of Medicine Solna Karolinska University Hospital and Karolinska Institutet Stockholm Sweden., Teeuwen L; Department of Clinical Immunology and Transfusion Medicine and Division of Immunology and Allergy, Department of Medicine Solna Karolinska University Hospital and Karolinska Institutet Stockholm Sweden., Czarnewski P; Science for Life Laboratory Department of Biochemistry and Biophysics National Bioinformatics Infrastructure Sweden Stockholm University Solna Sweden., Güclüler Akpinar G; Department of Clinical Immunology and Transfusion Medicine and Division of Immunology and Allergy, Department of Medicine Solna Karolinska University Hospital and Karolinska Institutet Stockholm Sweden., Sandberg A; Department of Oncology and Pathology Karolinska Institutet Science for Life Laboratory Solna Sweden., Cao X; Department of Oncology and Pathology Karolinska Institutet Science for Life Laboratory Solna Sweden., Pernemalm M; Department of Oncology and Pathology Karolinska Institutet Science for Life Laboratory Solna Sweden., Orre LM; Department of Oncology and Pathology Karolinska Institutet Science for Life Laboratory Solna Sweden., Gabrielsson S; Department of Clinical Immunology and Transfusion Medicine and Division of Immunology and Allergy, Department of Medicine Solna Karolinska University Hospital and Karolinska Institutet Stockholm Sweden., Eldh M; Department of Clinical Immunology and Transfusion Medicine and Division of Immunology and Allergy, Department of Medicine Solna Karolinska University Hospital and Karolinska Institutet Stockholm Sweden.
Jazyk: angličtina
Zdroj: Journal of extracellular vesicles [J Extracell Vesicles] 2021 Jul; Vol. 10 (9), pp. e12128. Date of Electronic Publication: 2021 Jul 22.
DOI: 10.1002/jev2.12128
Abstrakt: Extracellular vesicles (EVs) are increasingly tested as therapeutic vehicles and biomarkers, but still EV subtypes are not fully characterised. To isolate EVs with few co-isolated entities, a combination of methods is needed. However, this is time-consuming and requires large sample volumes, often not feasible in most clinical studies or in studies where small sample volumes are available. Therefore, we compared EVs rendered by five commonly used methods based on different principles from conditioned cell medium and 250 μl or 3 ml plasma, that is, precipitation (ExoQuick ULTRA), membrane affinity (exoEasy Maxi Kit), size-exclusion chromatography (qEVoriginal), iodixanol gradient (OptiPrep), and phosphatidylserine affinity (MagCapture). EVs were characterised by electron microscopy, Nanoparticle Tracking Analysis, Bioanalyzer, flow cytometry, and LC-MS/MS. The different methods yielded samples of different morphology, particle size, and proteomic profile. For the conditioned medium, Izon 35 isolated the highest number of EV proteins followed by exoEasy, which also isolated fewer non-EV proteins. For the plasma samples, exoEasy isolated a high number of EV proteins and few non-EV proteins, while Izon 70 isolated the most EV proteins. We conclude that no method is perfect for all studies, rather, different methods are suited depending on sample type and interest in EV subtype, in addition to sample volume and budget.
Competing Interests: Susanne Gabrielsson has a patent on B cell derived exosomes in immune therapy and is part of the Scientific Advisory Board of Anjarium Biosciences.
(© 2021 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)
Databáze: MEDLINE
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