Human neutrophil elastase (HNE) is a biomarker for chronic wounds and a therapeutic target for certain diseases. An unchecked influx of neutrophils, which contain about one pictogram of elastase per neutrophil, is responsible for degrading growth factors and collagen formation, indefinitely delaying the healing of chronic wounds. Nanocellulose when conjugated with HNE peptide substrates has the potential to improve point of care diagnostics used to assess the status of chronic wounds, by offering a higher transducer surface area for protease detection sensitivity. Over the years the use of cellulose and other polysaccharides as a solid support for peptides applicable to wound care has grown. Here cellulose nanocrystals and mirofibrillated cellulose (CNC) derived from cotton and wood respectively were compared as support surfaces, and were produced with varying material forms for attachment of elastase peptide substrates. Print cloth fabric, wood/cotton cellulose nanocrystals and microfibrillated cellulosic films were compared as putative transducer surfaces to host-peptide fluorescent sensors of HNE. Analytical approaches including attenuated total reflectance infrared spectroscopy (ATR-IR), ultraviolet visible spectroscopy (UV-VIS), mass spectrometry (MS), X-ray crystallography and elemental analysis (EA) were utilized to characterize the peptide-cellulose conjugates. The cellulose and nanocellulose-peptide conjugates were evaluated for their elastase detection activities using a microtiter enzyme assay to monitor fluorescence. Results of the relative sensitivity of HNE detection, surface area differences, degree of peptide substitution, and potential for applications to protease detection will be discussed.

Detection of human neutrophil elastase with fluorescent peptide sensors conjugated to nanocellulosic solid supports targeting wound care diagnostics

249th National Meeting (2015)

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