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Development of an enhanced proteomic method to detect prognostic and diagnostic markers of healing in chronic wound fluid

Fernandez, Melissa L., Broadbent, James A., Shooter, Gary K., Malda, Jos, & Upton, Zee (2008) Development of an enhanced proteomic method to detect prognostic and diagnostic markers of healing in chronic wound fluid. British Journal of Dermatology, 158(2), pp. 281-290.

Abstract

Background Chronic venous leg ulcers are a significant cause of pain, immobility and decreased quality of life for patients with these wounds. In view of this, research efforts are focused on multiple factors in the wound environment to obtain information regarding the healing of ulcers.

Objectives Chronic wound fluid (CWF), containing a complex mixture of proteins, is an important modulator of the wound environment, and therefore we hypothesized that these proteins may be indicators of the status of wounds and their potential to heal or otherwise. To explore this we developed and validated a proteomic approach to analyse CWF.

Methods In this study, pooled CWF was depleted of high abundant proteins using immunoaffinity chromatography. The flow-through and bound fractions were collected, concentrated, desalted and analysed using a range of techniques. Each fraction was further separated using two-dimensional (2D) gel electrophoresis and 2D liquid chromatography and analysed using mass spectrometry (MS).

Results Western blot analysis against three high abundant proteins confirmed the selective removal of these proteins from CWF. Critically, one-dimensional and 2D gel electrophoresis indicated that subsequent removal of these proteins enhanced the ability to detect proteins in low abundance in CWF. Further, MS demonstrated that depletion of these abundant proteins increased the detection of other proteins in these samples.

Conclusions Results obtained indicate that this approach significantly improves separation of proteins present in low concentrations in CWF. This will facilitate the identification of biomarkers in samples collected from patients with ulcers and lead to improved patient therapies and wound care approaches.

Impact and interest:

18 citations in Scopus
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16 citations in Web of Science®

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ID Code: 12981
Item Type: Journal Article
Additional Information: For more information, please refer to the journal's website (see hypertext link) or contact the author.
Additional URLs:
Keywords: wounds, wound healing, chronic wounds, cells and tissue, ihbi, tissue repair and regeneration, tissue repair, wound fluid, cwf, chronic wound fluid
ISSN: 0007-0963
Subjects: Australian and New Zealand Standard Research Classification > MEDICAL AND HEALTH SCIENCES (110000) > MEDICAL PHYSIOLOGY (111600) > Cell Physiology (111601)
Australian and New Zealand Standard Research Classification > MEDICAL AND HEALTH SCIENCES (110000) > MEDICAL MICROBIOLOGY (110800) > Medical Microbiology not elsewhere classified (110899)
Divisions: Past > QUT Faculties & Divisions > Faculty of Science and Technology
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: Copyright 2008 Blackwell Publishing
Copyright Statement: The definitive version is available at www.blackwell-synergy.com
Deposited On: 17 Mar 2008
Last Modified: 29 Feb 2012 23:39

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