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

Journal of Extracellular Vesicles

Patrick Pauwels¹  Geert Baggerman²  Hanny Willems²  Kurt Boonen²  An Jacobs²  Inge Mertens²  Eline Oeyen²  Kurt Van Mol³  William C Cho⁴  Karin Schildermans⁵  Christian Rolfo⁵ 

[1] Antwerp University Hospital (UZA);Flemish Institute for Technological Research (VITO), Mol;Pharmafluidics;Queen Elizabeth Hospital;University of Antwerp;

关键词: Extracellular vesicles;    small extracellular vesicles;    exosomes;    urine;    isolation;    size exclusion chromatography;    asymmetrical-flow field-flow fractionation;   

DOI  :  10.1080/20013078.2018.1490143

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown