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Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine

Extracellular vesicles (EVs) have a great potential in clinical applications. However, their isolation from different bodily fluids and their characterisation are currently not optimal or standardised. Here, we report the results of examining the performance of ultrafiltration combined with size exc...

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Autores principales: Oeyen, Eline, Van Mol, Kurt, Baggerman, Geert, Willems, Hanny, Boonen, Kurt, Rolfo, Christian, Pauwels, Patrick, Jacobs, An, Schildermans, Karin, Cho, William C, Mertens, Inge
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6032024/
https://www.ncbi.nlm.nih.gov/pubmed/29988836
http://dx.doi.org/10.1080/20013078.2018.1490143
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author Oeyen, Eline
Van Mol, Kurt
Baggerman, Geert
Willems, Hanny
Boonen, Kurt
Rolfo, Christian
Pauwels, Patrick
Jacobs, An
Schildermans, Karin
Cho, William C
Mertens, Inge
author_facet Oeyen, Eline
Van Mol, Kurt
Baggerman, Geert
Willems, Hanny
Boonen, Kurt
Rolfo, Christian
Pauwels, Patrick
Jacobs, An
Schildermans, Karin
Cho, William C
Mertens, Inge
author_sort Oeyen, Eline
collection PubMed
description Extracellular vesicles (EVs) have a great potential in clinical applications. However, their isolation from different bodily fluids and their characterisation are currently not optimal or standardised. Here, we report the results of examining the performance of ultrafiltration combined with size exclusion chromatography (UF-SEC) to isolate EVs from urine. The results reveal that UF-SEC is an efficient method and provides high purity. Furthermore, we introduce asymmetrical-flow field-flow fractionation coupled with a UV detector and multi-angle light-scattering detector (AF4/UV-MALS) as a characterisation method and compare it with current methods. We demonstrate that AF4/UV-MALS is a straightforward and reproducible method for determining size, amount and purity of isolated urinary EVs.
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spelling pubmed-60320242018-07-09 Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine Oeyen, Eline Van Mol, Kurt Baggerman, Geert Willems, Hanny Boonen, Kurt Rolfo, Christian Pauwels, Patrick Jacobs, An Schildermans, Karin Cho, William C Mertens, Inge J Extracell Vesicles Research Article Extracellular vesicles (EVs) have a great potential in clinical applications. However, their isolation from different bodily fluids and their characterisation are currently not optimal or standardised. Here, we report the results of examining the performance of ultrafiltration combined with size exclusion chromatography (UF-SEC) to isolate EVs from urine. The results reveal that UF-SEC is an efficient method and provides high purity. Furthermore, we introduce asymmetrical-flow field-flow fractionation coupled with a UV detector and multi-angle light-scattering detector (AF4/UV-MALS) as a characterisation method and compare it with current methods. We demonstrate that AF4/UV-MALS is a straightforward and reproducible method for determining size, amount and purity of isolated urinary EVs. Taylor & Francis 2018-07-03 /pmc/articles/PMC6032024/ /pubmed/29988836 http://dx.doi.org/10.1080/20013078.2018.1490143 Text en © 2018 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Oeyen, Eline
Van Mol, Kurt
Baggerman, Geert
Willems, Hanny
Boonen, Kurt
Rolfo, Christian
Pauwels, Patrick
Jacobs, An
Schildermans, Karin
Cho, William C
Mertens, Inge
Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine
title Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine
title_full Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine
title_fullStr Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine
title_full_unstemmed Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine
title_short Ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine
title_sort ultrafiltration and size exclusion chromatography combined with asymmetrical-flow field-flow fractionation for the isolation and characterisation of extracellular vesicles from urine
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6032024/
https://www.ncbi.nlm.nih.gov/pubmed/29988836
http://dx.doi.org/10.1080/20013078.2018.1490143
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