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CYP450 2B4 covalently attached to carbon and gold screen printed electrodes by diazonium salt and thiols monolayers
Authors:M.A. Alonso-Lomillo,C. Yardimci,M.J. Arcos-Martí  nez
Affiliation:a Analytical Chemistry Department, Faculty of Sciences, University of Burgos, Pza. Misael Bañuelos s/n, 09001 Burgos, Spain
b Analytical Chemistry Department, Faculty of Pharmacy, Hacettepe University, 06100 Sihhiye, Ankara, Turkey
Abstract:An easy covalent immobilization method used to develop enzyme biosensors based on carbon and gold screen printed electrodes (SPCEs and gold SPEs) is described. The linkage of biomolecules through 4-nitrobenzenediazonium tetrafluoroborate, mercaptopropionic acid and thioctic acid monolayers has been attempted using bare SPCEs and gold SPEs, as well as gold nanoparticles (AuNPs) modified SPCEs and gold SPEs. Direct covalent attachment of Cytochrome P450 2B4 (CYP450 2B4) to the transducer has been carried out by carbodiimide and hydroxysuccinimide. Experimental variables in the immobilization process and in the chronoamperometric determination of Phenobarbital (PB) have been optimized by the experimental design methodology. Reproducibility of the different biosensors has been checked under the optimum conditions, yielding values lower than 6%. Their performances have been shown by the determination of PB in pharmaceutical drugs.
Keywords:Screen printed electrodes   Gold nanoparticles   Diazonium salt   Self-assembled monolayer   Biosensor
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