Human neutrophil elastase peptide sensors conjugated to cellulosic and nanocellulosic materials: part I, synthesis and characterization of fluorescent analogs

Authors: Edwards, Judson - Vince; Fontenot, Krystal; HALDANE, DAVID - Innovatech-Engineering; Prevost, Nicolette; Condon, Brian; Grimm, Casey

Submitted to: Cellulose
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/28/2016
Publication Date: 3/26/2016
Citation: Edwards, J.V., Fontenot, K.R., Haldane, D., Prevost, N.T., Condon, B.D., Grimm, C.C. 2016. Human neutrophil elastase peptide sensors conjugated to cellulosic and nanocellulosic materials: part I, synthesis and characterization of fluorescent analogs. Cellulose. 23(2):1283-1295.

Interpretive Summary: A natural source of cotton and wood is employed to generate cellulose and nanocellulose sensors for the detection of human neutrophil elastase that is not only a marker for chronic wounds but is also a marker for inflammatory diseases. To produce the sensors cellulose (cotton filter paper and print cloth fabric) and nanocellulose (wood cellulose nanocrystals and wood nanocellulose composites) transducers were covalently attached to a fluorogenic elastase peptide substrate, which detects human neutrophil elastase. Characterization results support the transducers as a good surface for generating sensors. The study shows that the nanocellulose transducers, with emphasis on the wood cellulose nanocrystals, had the highest level of the peptide substrate immobilized on the surface and is a promising transducer surface for biosensors that detect human neutrophil elastase.

Technical Abstract: Here we describe the synthesis and characterization of peptide conjugated cellulose and nanocellulose materials as sensors for fluorescent detection of human neutrophil elastase (HNE). The cellulose sensor surfaces selected are filter paper (FP) and print cloth (PC) fabric, which are composed of processed cotton fibers. The nanocellulose based sensors have transducer surfaces comprised of cellulose nanocrystals (wCNC) and microfibrillated cellulose (MFC) derived from wood that are fabricated as films wood nanocellulose composities (wNCCs) consisting of blended quantities of nanocrystalline and microfibrillated cellulose at 66/33 and 50/50 ratios. The fluorescent peptide HNE substrate, succinamidyl-Ala-Ala-Pro-Val-amidylcoumadin (Pep) was attached to both cellulosic and nanocellulosic matrices and conjugated peptide analogs were confirmed by Mass Spectrometry and Infrared (IR). The nanocellulose biosensors wCNC-Pep (3c) and wNCC-Pep (4c, 66/33 and 5c, 50/50) have higher levels of peptide incorporation than the cellulosic biosensors FP-Pep (Ic) and PCPep (2c). The range of incorporation for the nanocellulose sensors is 30 - 80 ug/mg and for the cellulosic sensors 7 - 25 ug/mg. The degree of substitution of peptide was found to be in the order of approximate number of peptides per 200 anyhydroglucose residues, 1 in PC-Pep (2c), 2 in FP-Pep (Ic), 4 in wNCC-Pep (5c, 50/50), 6 in wNCC-Pep (4c, 66/33), and 12 in wCNC-Pep (3c). Of the cellulose and nanocellulose biosensors, the wCNC-Pep (3c) has the highest level of peptide incorporation, which maizes it the preferred sensor matrix for human neutrophil elastase.