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Sample collection in clinical proteomics—Proteolytic activity profile of serum and plasma
Article first published online: 11 MAY 2014
© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
PROTEOMICS - Clinical Applications
Special Issue: Proteases and Disease
Volume 8, Issue 5-6, pages 299–307, June 2014
How to Cite
Jambunathan, K. and Galande, A. K. (2014), Sample collection in clinical proteomics—Proteolytic activity profile of serum and plasma. Prot. Clin. Appl., 8: 299–307. doi: 10.1002/prca.201300037
- Issue published online: 10 JUN 2014
- Article first published online: 11 MAY 2014
- Accepted manuscript online: 10 APR 2014 05:56AM EST
- Manuscript Accepted: 3 APR 2014
- Manuscript Revised: 23 FEB 2014
- Manuscript Received: 13 MAY 2013
- SRI International's Center for Advanced Drug Research
- Combinatorial library;
- Fluorogenic probes;
Proteolytic enzymes are promising diagnostic targets since they play key roles in diverse physiological processes and have been implicated in numerous human diseases. Human blood is a relatively noninvasive source for disease-specific protease biomarker detection and subsequent translation into diagnostic tests. However, the choice of serum or plasma, and more specifically, which anticoagulant to choose in plasma preparation, is important to address in the sample preparation phase of biomarker discovery.
We have previously utilized a combinatorial library of internally quenched fluorogenic probes to successfully map the global proteolytic profiles of various biological fluids. In this study, we utilized the platform to ascertain the impact of three commonly used anticoagulants (EDTA, heparin, and citrate) on the proteolytic activity profile of plasma and serum collected from a healthy Caucasian male.
Serum and plasma citrate were observed to be most proteolytically active, followed by plasma heparin and then plasma EDTA. Detailed analysis of the amino acid distribution of motifs cleaved and not cleaved by the samples offered significant insights in to active proteolytic components within them.
Conclusion and clinical relevance
Broad quantitative comparison of proteolytic profiles of these samples revealed several novel insights related to the differential substrate recognition of proteases present in these biological fluids.