title: Immobilization-stabilization of proteins on nanofibrillated cellulose derivatives and their bioactive film formation.
abstract: In a number of different applications for enzymes and specific binding proteins a key technology is the immobilization of these proteins to different types of supports. In this work we describe a concept for protein immobilization that is based on nanofibrillated cellulose (NFC). NFC is a form of cellulose where fibers have been disintegrated into fibrils that are only a few nanometers in diameter and have a very large aspect ratio. Proteins were conjugated through three different strategies using amine, epoxy, and carboxylic acid functionalized NFC. The conjugation chemistries were chosen according to the reactive groups on the NFC derivatives; epoxy amination, heterobifunctional modification of amino groups, and EDC/s-NHS activation of carboxylic acid groups. The conjugation reactions were performed in solution and immobilization was performed by spin coating the protein-NCF conjugates. The structure of NFC was shown to be advantageous for both protein performance and stability. The use of NFC allows all covalent chemistry to be performed in solution, while the immobilization is achieved by a simple spin coating or spreading of the protein-NFC conjugates on a support. This allows more scalable methods and better control of conditions compared to the traditional methods that depend on surface reactions.
authors.affiliation: VTT , Technical Research Centre of Finland, Bio and Process Technology, Tietotie 2, P.O. Box 1000, FIN-02044 VTT, Finland. extsuvi.arola@vtt.fi
authors.forename: Suvi
authors.initials: S
authors.lastname: Arola
authors.forename: Tekla
authors.initials: T
authors.lastname: Tammelin
authors.forename: Harri
authors.initials: H
authors.lastname: Setälä
authors.forename: Antti
authors.initials: A
authors.lastname: Tullila
authors.forename: Markus B
authors.initials: MB
authors.lastname: Linder
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry* / Q000378:metabolism; D000588:Amines / Q000737:chemistry; D000818:Animals; D002264:Carboxylic Acids / Q000737:chemistry; D002417:Cattle; D002482:Cellulose / Q000737:chemistry*; D004853:Epoxy Resins / Q000737:chemistry; D006854:Hydrocortisone / Q000276:immunology / Q000378:metabolism; D055767:Immobilized Proteins / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D053758:Nanoparticles / Q000737:chemistry*; D011134:Polysaccharides / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000588:Amines; D002264:Carboxylic Acids; D004853:Epoxy Resins; D055767:Immobilized Proteins; D011134:Polysaccharides; D012710:Serum Albumin, Bovine; D002482:Cellulose; D000469:Alkaline Phosphatase; D006854:Hydrocortisone