Ready-made chromatography columns for extracellular vesicle isolation from plasma.
Welton, Joanne Louise
Webber, Jason Paul
MetadataDangos cofnod eitem llawn
Proteomic 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.
Journal of Extracellular Vesicles;
Welton, 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.
Dynodwr Gwrthrych Digidol (DOI)http://dx.doi.org/10.3402/jev.v4.27269
HawliauThe article is made available in this repository under the Creative Commons Attribution Non-Commercial 3.0 Unported License
Cancer Research Wales, Prostate Cancer UK, Movember, MS Society
Yn dangos eitemau sy’n perthyn drwy deitl, awdur, pwnc a chrynodeb.
Proteomics analysis of vesicles isolated from plasma and urine of prostate cancer patients using a multiplex, aptamer-based protein array Welton, Joanne Louise; Brennan, Paul; Gurney, Mark; Webber, Jason Paul; Spray, Lisa Kate; Carton, David Gil; Falcon-Perez, Juan Manuel; Walton, Sean Peter; Mason, Malcolm David; Tabi, Zsuzsanna; Clayton, Aled (Co-Action, 2016-06-29)Proteomics analysis of biofluid-derived vesicles holds enormous potential for discovering non-invasive disease markers. Obtaining vesicles of sufficient quality and quantity for profiling studies has, however, been a major ...
Connolly, Katherine; Wadey, Rebecca; Matthew, Donna; Johnson, Errin; Rees, D. Aled; James, Philip (Oxford University Press, 2018-07-16)Adipocyte-derived extracellular vesicles (EVs) may serve as novel endocrine mediators of adipose tissue and impact upon vascular health. However, it is unclear whether adipocyte-derived EVs are present in the human ...
Cerebrospinal fluid extracellular vesicle enrichment for protein biomarker discovery in neurological disease; multiple sclerosis Welton, Joanne Louise; Loveless, Samantha; Stone, Timothy; von Ruhland, Chris; Robertson, Neil; Clayton, Aled (Taylor & Francis, 2017-09-03)The discovery of disease biomarkers, along with the use of “liquid biopsies” as a minimally invasive source of biomarkers, continues to be of great interest. In inflammatory diseases of the central nervous system (CNS), ...