ABSTRACT
We report a rapid and versatile procedure for the preparation of photoreactive polymers and light-induced immobilization of proteins onto such polymers. Photoreactive controlled-pore glass, silica gel, glass slide, and polystyrene microtiter plate are prepared in 40-60s by microwave irradiation of the respective amino polymers and 1-fluoro-2-nitro-4-azidobenzene. Azido group, now part of the polymer, yields highly reactive nitrene under ultraviolet (UV) light at 365 nm. Thus, when photoreactive polymer and horseradish peroxidase or glucose oxidase are exposed to UV light, the reactive nitrene immobilizes the protein molecules in 10 to 20 min through covalent bonding. As nitrene has a property of inserting into C-H bond, the method may find potential applications for immobilization of biomolecules irrespective of their functional groups.
Subject(s)
Azides/chemistry , Enzymes, Immobilized/chemistry , Microwaves , Photochemistry/methods , Polymers/chemical synthesis , Polymers/radiation effects , Glass/chemistry , Glucose Oxidase/chemistry , Glucose Oxidase/radiation effects , Horseradish Peroxidase/chemistry , Horseradish Peroxidase/radiation effects , Nitrobenzenes/chemistry , Silica Gel , Silicon Dioxide/chemistry , Ultraviolet RaysABSTRACT
Polystyrene, polypropylene, and polyethylene surfaces were activated by exposing 1-fluoro-2-nitro-4-azidobenzene coated surface to sunlight. Sunlight intensity of 26,300 lux was found optimum beyond which no appreciable increase in activation was observed. Five-minutes sunlight exposure gave better activated surface than 5 min 365-nm UV light exposure. The efficacy of sunlight-mediated activated surfaces was demonstrated by covalently immobilizing proteins onto them. Horseradish peroxidase when immobilized onto the sunlight-activated surfaces showed more than twofold increase in immobilization than the surface without activation. Thus, sunlight being a versatile, eco-friendly, and clean energy source can be a potential alternative for activation of inert surface for covalent attachment of biomolecule such as protein, DNA, or carbohydrate.
Subject(s)
Azides/radiation effects , Enzymes, Immobilized/chemistry , Nitrobenzenes/radiation effects , Photochemistry/methods , Plastics/chemistry , Sunlight , Azides/chemistry , Horseradish Peroxidase/chemistry , Nitrobenzenes/chemistry , Proteins/chemistry , Ultraviolet RaysABSTRACT
Polypropylene and polyethylene surfaces are activated by introducing an active functional group through 1-fluoro-2 nitro-4-azidobenzene by UV irradiation. Horseradish peroxidase and glucose oxidase are immobilized onto the activated surfaces, simply by incubating the enzymes at 37 degrees C. When untreated surfaces are used, insignificant immobilization of the enzymes is observed.
