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

doi:10.1080/20013078.2018.1490143

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

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

Keywords:

Science & Technology, Life Sciences & Biomedicine, Cell Biology, Extracellular vesicles, small extracellular vesicles, exosomes, urine, isolation, size exclusion chromatography, asymmetrical-flow field-flow fractionation, NANOPARTICLE TRACKING ANALYSIS, MASS-SPECTROMETRY, PROTEOMIC ANALYSIS, EXOSOMES, MICROPARTICLES, MICROVESICLES, STORAGE, 0601 Biochemistry and Cell Biology, 3101 Biochemistry and cell biology

Abstract:

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.