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 ORCID: https://orcid.org/0000-0001-8792-0499, Gurney, Mark ORCID: https://orcid.org/0000-0003-1119-6638, Webber, Jason Paul ORCID: https://orcid.org/0000-0003-4772-3014, Spary, Lisa Kate, Gil Carton, David, Falcón-Pérez, Juan Manuel, Walton, Sean Peter, Mason, Malcolm David ORCID: https://orcid.org/0000-0003-1505-2869, Tabi, Zsuzsanna and Clayton, Aled ORCID: https://orcid.org/0000-0002-3087-9226 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. 10.3402/jev.v5.31209

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Abstract

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.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Medicine
Subjects:	R Medicine > R Medicine (General)
Uncontrolled Keywords:	plasma, urine prostate, proteomics, protein array
Publisher:	Co-Action Publishing
ISSN:	2001-3078
Date of First Compliant Deposit:	21 September 2016
Date of Acceptance:	17 April 2016
Last Modified:	12 Nov 2024 08:30
URI:	https://orca.cardiff.ac.uk/id/eprint/94709

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