Hepburn, S, Cairns, DA, Jackson, D et al. (7 more authors) (2015) An analysis of the impact of pre-analytical factors on the urine proteome: Sample processing time, temperature, and proteolysis. PROTEOMICS - Clinical Applications, 9 (5-6). pp. 507-521. ISSN 1862-8346
Abstract
Purpose: We have examined the impact of sample processing time delay, temperature, and the addition of protease inhibitors (PIs) on the urinary proteome and peptidome, an important aspect of biomarker studies. Experimental design: Ten urine samples from patients with varying pathologies were each divided and PIs added to one-half, with aliquots of each then processed and frozen immediately, or after a delay of 6 h at 4°C or room temperature (20–22°C), effectively yielding 60 samples in total. Samples were then analyzed by 2D-PAGE, SELDI-TOF-MS, and immunoassay. Results: Interindividual variability in profiles was the dominant feature in all analyses. Minimal changes were observed by 2D-PAGE as a result of delay in processing, temperature, or PIs and no changes were seen in IgG, albumin, β2-microglobulin, or α1-microglobulin measured by immunoassay. Analysis of peptides showed clustering of some samples by presence/absence of PIs but the extent was very patient-dependent with most samples showing minimal effects. Conclusions and clinical relevance: The extent of processing-induced changes and the benefit of PI addition are patient- and sample-dependent. A consistent processing methodology is essential within a study to avoid any confounding of the results.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2014 The Authors PROTEOMICS Clinical Applications Published by Wiley-VCH Verlag GmbH & Co. KGaA www.clinical.proteomics-journal.com. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
Keywords: | Peptides; Protease inhibitor; Proteomics; Temperature; Urine |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) > Inst of Clinical Trials Research (LICTR) (Leeds) |
Funding Information: | Funder Grant number MRC G0802416 |
Depositing User: | Symplectic Publications |
Date Deposited: | 13 May 2016 14:26 |
Last Modified: | 13 May 2016 14:26 |
Published Version: | http://doi.org/10.1002/prca.201400079 |
Status: | Published |
Publisher: | Wiley-VCH Verlag |
Identification Number: | 10.1002/prca.201400079 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:92494 |