Synthesis and characterization of TEMPO-oxidized peptide-cellulose conjugate biosensors for detecting human neutrophil elastase

Here we describe the synthesis and characterization of a peptide-cellulose conjugate biosensor based on TEMPO-oxidized nanofibrillated cellulose (tNFC) for detecting elevated levels of human neutrophil elastase (HNE) in chronic wounds. The fluorescent peptide HNE substrate constructed from n-succiny...

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Bibliographic Details
Published in:Cellulose (London) 2022, Vol.29 (2), p.1293-1305
Main Authors: Mackin, Robert T., Fontenot, Krystal R., Edwards, J. Vincent, Prevost, Nicolette T., Grimm, Casey, Condon, Brian D., Liebner, Falk, Jordan, Jacobs H., Easson, Michael W., French, Alfred D.
Format: Article
Language:English
Subjects:
Aerogels
Bioorganic Chemistry
Biosensors
Cellulose
Ceramics
Chemistry
Chemistry and Materials Science
Composites
Conjugates
Coumarin
Crystallites
Elastase
Fluorescence
Glass
Hydrogels
Natural Materials
Neutrophils
Organic Chemistry
Original Research
Peptides
Physical Chemistry
Polyethylene glycol
Polymer Sciences
Substrates
Sustainable Development
Synthesis
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Summary:Here we describe the synthesis and characterization of a peptide-cellulose conjugate biosensor based on TEMPO-oxidized nanofibrillated cellulose (tNFC) for detecting elevated levels of human neutrophil elastase (HNE) in chronic wounds. The fluorescent peptide HNE substrate constructed from n-succinyl-Ala-Pro-Ala-7-amino-4-methyl-coumarin was attached to the TEMPO-oxidized cellulose surface via polyethylene glycol linker. The characterization of the biosensor conjugate shows a high degree of peptide incorporation onto the surface with the degree of substitution of 0.057. The relatively small crystallite size of 26.0 Å compared to other cellulose- and nanocellulose-based materials leads to a large specific surface area which can promote access of HNE to the enzyme substrates due to decreased steric interactions. Likewise, the porosity for tNFC was found to be higher than all other samples, including the nanocellulosic aerogel, lending to its hydrogel-like nature. The properties of tNFC were compared to other cellulose-based materials. The volume of each crystallite and volume ratio to the largest sample was calculated. tNFC was found to occupy the smallest space resulting in high amounts of sensors per crystallite unit volume. With a small crystallite volume and large number of sensors, the tNFC peptide-cellulose conjugate biosensor could provide a more sensitive system and is a good candidate for point of care diagnostic devices for detecting elevated protease levels in humans.
ISSN:0969-0239
1572-882X
DOI:10.1007/s10570-021-04362-z