Mr Tuo Zang1,2,3, Dr Leila Cuttle1,2,4, Dr Daniel Broszczak1,2,5, Dr James Broadbent1,2,6, Ms Catherine Tanzer1,2,4, Dr Tony Parker1,2
1Institute of Health and Biomedical Innovation, Kelvin Grove, Australia, 2Queensland University Of Technology, Kelvin Grove, Australia, 3Wound Management Innovation Co-operative Research Centre, Brisbane, Australia, 4Institute of Health and Biomedical Innovation at the Centre for Children’s Health Research, South Brisbane, Australia, 5Australian Catholic University, Banyo, Australia, 6Sciex P/L, Brisbane, Australia
Blister fluid (BF) is a sample matrix that can reflect both systemic and local microenvironment responses to burn injury. Our aim was to investigate the proteome of burn blister fluid using liquid chromatography tandem mass spectrometry (LC-MS/MS), which provides an objective and quantitative measurement of biomolecules in the wound site. This enables a non-invasive investigation of the cutaneous biological response to different burn severities, which may aid in the clinical diagnosis and monitor the response of treatment.
In order to generate a comprehensive peptide spectral library for quantitative information acquisition, a subset of BF samples were pooled according to burn depth (12 superficial, 12 deep-partial thickness, and 4 full thickness) and fractionated by four different methods, prior to LC-MS/MS analysis. Subsequently, all individual BF samples (n=87) were analysed using LC-MS/MS in quantitative mode (SWATH) to obtain the protein abundance. More than 800 individual proteins were identified and formed the basis of a BF protein library. Finally, more than 600 proteins in each individual sample was quantitatively profiled. This proteomic data was correlated with different clinical parameters, such as burn depth, time to re-epithelialisation, first aid, and burn size.
The statistical analysis of these data revealed the key biochemical differences that stratify sub-groups within the clinically relevant parameters. We have shown that the blister fluid proteome could be used to assist the classification of paediatric burn wounds by different burn depths and prognose time to re-epithelialisation at an earlier stage. Interestingly differences in the application of first aid may also be reflected in the burn wound protein profile. This may help deepen our understanding of the biological effect of first aid treatment.
Tuo Zang is a PhD student of the Tissue Repair and Translational Physiology Program (TRTP) at the Institute of Health and Biomedical Innovation (IHBI) at the Queensland University of Technology. His research has focused on proteomics and metabolomics, especially quantitative proteomics analysis of clinical samples. His PhD study characterised blister fluid from paediatric burn injury using LC-MS/MS based proteomics and NMR based metabolomics.