Photoreactive cellulose membrane--A novel matrix for covalent immobilization of biomolecules.
We report a simple and mild procedure for the preparation of a photoreactive cellulose membrane capable of forming a covalent bond with a biomolecule in presence of 365 nm UV light. Photoreactive cellulose membrane was prepared by the reaction of fluoro group of 1-fluoro-2-nitro-4-azidobenzene (FNAB) and hydroxyl group of the cellulose in an alkaline medium. X-ray photoelectron spectroscopy (XPS) of the photoreactive cellulose confirmed the incorporation of FNAB moiety. Azido group of the photoreactive membrane on exposure to UV light transforms into highly reactive nitrene which binds with a protein. The efficacy of the activated membrane was checked by immobilizing glucose oxidase (GOD) onto it in presence of light. Immobilized GOD was found to have improved thermal, pH and storage stability. Photoreactive cellulose membrane was successfully used in enzyme-linked immunosorbent assay (ELISA) technique. The antibody immobilized onto such support by UV irradiation in 30 min showed similar ELISA value than the antibody immobilized onto a polystyrene ELISA plate in 12h incubation at 4 degrees C by conventional method.
Bora
Utpal
U
Institute of Genomics and Integrative Biology (CSIR), Mall Road, Delhi-110007, India.
Sharma
Pragya
P
Kannan
Krishnamoorthy
K
Nahar
Pradip
P
D000327:Adsorption; D001704:Biopolymers / Q000032:analysis / Q000737:chemistry*; D002482:Cellulose / Q000737:chemistry* / Q000528:radiation effects*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D004800:Enzymes, Immobilized / Q000032:analysis / Q000737:chemistry*; D008422:Materials Testing; D008567:Membranes, Artificial; D010777:Photochemistry / Q000379:methods*; D014466:Ultraviolet Rays
D001704:Biopolymers; D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D002482:Cellulose