Burn injury is a common and traumatic event in the paediatric population. At present, the diagnosis of burn injury severity is largely dependent on the clinician’s experience. A better understanding of the biochemistry of burn injury would assist with the development of objective and quantitative measures to aid diagnosis. Burn blister fluid (BF) is considered to be a viable source of biomolecules that reflect relevant systemic responses and the local microenvironment.
In order to generate a comprehensive peptide spectral library, a subset of BF samples were pooled according to burn depth (12 superficial (S), 12 deep-partial thickness (D), and 4 full thickness (F)) and fractionated by four different methods, including ultrafiltration, SDS-PAGE, OFFGel isoelectric focusing and immuno-depletion, prior to digestion and subsequent LC-MS/MS analysis in data-dependent acquisition mode. All individual BF samples (n=100) were then analysed using LC-MS/MS in data independent acquisition mode (SWATH) to obtain quantitative data.
More than 800 individual proteins were identified and formed the basis of a BF peptide spectral library. The relative ion abundance of more than 600 proteins in every individual sample was extracted and correlated with different clinical parameters, such as burn depth, time to re-epithelialisation, first aid treatment, and burn size. Analysis of these data using orthogonal partial least squares–discriminant analysis (OPLS-DA) revealed the key biochemical differences that stratify sub-groups within the clinically relevant parameters..
We have shown that the blister fluid proteome can be used to classify paediatric burn wounds by different burn depths and by other clinically relevant parameters. These markers are under further investigation to determine their viability as clinic