Title: Nanocellulose-based biosensors: design, preparation, and activity of peptide-linked cotton cellulose nanocrystals having fluorimetric and colorimetric elastase detection sensitivity Authors
Submitted to: Engineering
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: May 16, 2013
Publication Date: September 20, 2013
Citation: Edwards, J.V., Prevost, N.T., French, A.D., Concha, M.C., Delucca, A., Wu, Q. 2013. Nanocellulose-based biosensors: design, preparation, and activity of peptide-linked cotton cellulose nanocrystals having fluorimetric and colorimetric elastase detection sensitivity. Engineering. 5:20-28. Interpretive Summary: Biosensors may be defined as the interface of a molecular or receptor-recognition property with a cellular or biochemical activity, triggering a ‘bio-molecular switch’ that in turn is connected with a detector signal. Advances in biosensor technology hold promise to revolutionize healthcare and diagnosis, through assessment of health status, disease onset, and progression with non-invasive methods. This is especially the case in the area of sensor and imaging for wound healing, where approaches for biochemical and cellular markers are emerging. This paper outlines the design, preparation and activity of cellulose based biosensors, and in particular discusses a biosensor based on cotton cellulose nanocrystals for their ability to be used in combination with peptides and proteins to colorimetrically and fluorimetrically detect small concentrations of proteases. This new technology has promise to be used in point of care diagnostic detection of harmful proteases. Thus, the manuscript discusses an approach to design, preparing and assessing the activity of CCN peptides to detect human neutrophil elastase at levels found in chronic wounds, and an approach to using the technology as a biosensor.
Technical Abstract: Nanocrystalline cellulose is an amphiphilic, high surface area material that can be easily functionalized and is biocom-patible and eco-friendly. It has been used singularly and in combination with other nanomaterials to optimize biosensor design. The attachment of peptides and proteins to nanocrystalline cellulose and their proven retention of activity pro- vide a route to bioactive conjugates useful in designs for point of care biosensors. Elastase is a biomarker for a number of inflammatory diseases including chronic wounds, and its rapid sensitive detection with a facile approach to sensing is of interest. An increased interest in the use of elastase sensors for point of care diagnosis is resulting in a variety of ap- proaches to elsastase sensors utilizing different detection technologies. Here elastase substrate peptide-celluose conju- gates synthesized as colorimetric and fluorescent sensors on cotton cellulose nanocrystals are compared. The structure of the sensor peptide-nanocellulose crystals when modeled with computational crystal structure parameters demon- strates the spatio-stoichiometric features of the nanocrystalline surface that allows ligand to active site protease interact- tion. An understanding of the structure/function relations of enzyme and conjugate substrate of the peptides covalently attached to nancellulose has implications for enhancing the biomolecular transducer. The potential applications of both fluorescent and colorimetric detection to markers like elastase using peptide cotton cellulose nanocrystals as a trans- ducer surface to model point of care biosensors for protease detection are discussed.