Proteomics analysis of vesicles isolated from plasma and urine of prostate cancer patients using a multiplex, aptamer-based protein array
Welton, Joanne Louise
Webber, Jason Paul
Spray, Lisa Kate
Carton, David Gil
Falcon-Perez, Juan Manuel
Walton, Sean Peter
Mason, Malcolm David
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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 problem, as samples are often replete with co-isolated material that can interfere with the identification of genuine low abundance, vesicle components. Here, we used a combination of ultracentrifugation and size-exclusion chromatography to isolate and analyse vesicles of plasma or urine origin. We describe a sample-handling workflow that gives reproducible, quality vesicle isolations sufficient for subsequent protein profiling. Using a semi-quantitative aptamer-based protein array, we identified around 1,000 proteins, of which almost 400 were present at comparable quantities in plasma versus urine vesicles. Significant differences were, however, apparent with elements like HSP90, integrin αVβ5 and Contactin-1 more prevalent in urinary vesicles, while hepatocyte growth factor activator, prostate-specific antigen–antichymotrypsin complex and many others were more abundant in plasma vesicles. This was also applied to a small set of specimens collected from men with metastatic prostate cancer, highlighting several proteins with the potential to indicate treatment refractory disease. The study provides a practical platform for furthering protein profiling of vesicles in prostate cancer, and, hopefully, many other disease scenarios.
Journal of Extracellular Vesicles
Welton, J.L., Brennan, P., Gurney, M., Webber, J.P., Spray, L.K., Carton, D.G., Falcon-Perez, J.M., Walton, S.P., Mason, M.D., Tabi, Z. and Clayton, A. (2016) 'Proteomics analysis of vesicles isolated from plasma and urine of prostate cancer patients using a multiplex, aptamer-based protein array', Journal of Extracellular Vesicles, 5, 31209
This article was published in Journal of Extracellular Vesicles on 29 June 2016 (online), available open access at http://dx.doi.org/10.3402/jev.v5.31209
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