Open AccessUltrafiltration and size exclusion chromatography combined with asymmetrical‐flow field‐flow fractionation for the isolation and characterisation of extracellular vesicles from urine
Author(s) -
Oeyen Eline,
Van Mol Kurt,
Baggerman Geert,
Willems Hanny,
Boonen Kurt,
Rolfo Christian,
Pauwels Patrick,
Jacobs An,
Schildermans Karin,
Cho William C,
Mertens Inge
Publication year - 2018
Publication title -
journal of extracellular vesicles
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.94
H-Index - 68
ISSN - 2001-3078
DOI - 10.1080/20013078.2018.1490143
Subject(s) - ultrafiltration (renal) , extracellular vesicles , chromatography , size exclusion chromatography , fractionation , field flow fractionation , multiangle light scattering , chemistry , isolation (microbiology) , nanoparticle tracking analysis , materials science , light scattering , microvesicles , scattering , bioinformatics , biology , biochemistry , optics , physics , enzyme , microbiology and biotechnology , microrna , gene
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.