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Immobilization of monoamine oxidase on eggshell membrane and its application in designing an amperometric biosensor for dopamine
Authors:Preeti Joshi  Harish C Joshi  Sunil Kumar Sanghi  Subir Kundu
Institution:1. School of Biochemical Engineering, Institute of Technology, Banars Hindu University, Varanasi, India, 221005
3. IBM India Pvt. Ltd., Pune, India, 412308
2. Microfluidics & MEMS Centre, Advanced Materials and Processes Research Institute, Bhopal, India, 462026
Abstract:An amperometric biosensor for dopamine is described. It is based on the enzyme monoamine oxidase immobilized on a glutaraldehyde-activated eggshell membrane that was deposited on a glassy carbon electrode. The Michaelis-Menten constant (Km) is 0.087 mM. Optimum pH and temperature conditions were obtained at pH 7.0 and 37 °C, respectively. The sensor showed a detection limit of 20 μM, a linear range from 50 μM to 250 μM, and a storage stability of ~25 days. In order to further improve the performance, a Nafion coating was applied on the electrode surface which gave favorable results with respect to shelf life of the enzyme (~40 days), the limit of detection, and the selectivity over ascorbic acid and uric acid.
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