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dc.contributor.authorWelton, Joanne Louise
dc.contributor.authorWebber, Jason Paul
dc.contributor.authorBotos, Laur-Alexandru
dc.contributor.authorJones, Michael
dc.contributor.authorClayton, Aled
dc.date.accessioned2016-02-01T14:20:51Z
dc.date.available2016-02-01T14:20:51Z
dc.date.issued2015
dc.identifier.citationWelton, J.L., Webber, J.P., Botos, L.A., Jones, M. and Clayton, A. (2015) 'Ready-made chromatography columns for extracellular vesicle isolation from plasma', Journal of Extracell Vesicles, 4:27269.en_US
dc.identifier.issn2001-3078
dc.identifier.urihttp://hdl.handle.net/10369/7609
dc.description.abstractProteomic studies of circulating vesicles are hampered by difficulties in purifying vesicles from plasma and serum. Isolations are contaminated with high-abundance blood proteins that may mask genuine vesicular-associated proteins and/or simply provide misleading data. In this brief report, we explored the potential utility of a commercially available size exclusion chromatography column for rapid vesicle purification. We evaluated the performance of the column, with cancer cell line conditioned medium or healthy donor plasma, in terms of removing non-vesicular protein and enriching for vesicles exhibiting exosome characteristics. Serial fractions revealed a peak for typical exosomal proteins (CD9, CD81 etc.) that preceded the peak for highly abundant proteins, including albumin, for either sample type, and harvesting only this peak would represent elimination of >95% of protein from the sample. The columns showed good reproducibility, and streamlining the workflow would allow the exosome-relevant material to be collected in less than 10 minutes. Surprisingly, however, subsequent post-column vesicle concentration steps whilst resulting in some protein loss also lead to low vesicle recoveries, with a net effect of reducing sample purity (assessed by the particle-to-protein ratio). The columns provide a convenient, reproducible and highly effective means of eliminating >95% of non-vesicular protein from biological fluid samples such as plasma.en_US
dc.description.sponsorshipCancer Research Wales, Prostate Cancer UK, Movember, MS Societyen_US
dc.language.isoenen_US
dc.publisherCo-action Publishingen_US
dc.relation.ispartofseriesJournal of Extracellular Vesicles;
dc.rightsThe article is made available in this repository under the Creative Commons Attribution Non-Commercial 3.0 Unported License
dc.subjectplasmaen_US
dc.subjectexosomesen_US
dc.titleReady-made chromatography columns for extracellular vesicle isolation from plasma.en_US
dc.typeArticleen_US
dc.identifier.doihttp://dx.doi.org/10.3402/jev.v4.27269
dc.date.dateAccepted2015-03-04


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