Uricase-adsorbed carbon-felt reactor coupled with a peroxidase-modified carbon-felt-based H2O2 detector for highly sensitive amperometric flow determination of uric acid.
ABSTRACT Uricase (urate oxidase, UOx) was adsorbed onto a porous carbon-felt (CF) surface and the resulting UOx-adsorbed CF (UOx-CF) was successfully used as a column-type enzyme reactor coupled with a peroxidase-adsorbed CF-based bioelectrocatalytic H(2)O(2) flow-detector to fabricate a flow-amperometric biosensor for uric acid. In this flow-biosensor system, H(2)O(2) produced in the UOx-CF reactor was cathodically detected by horseradish peroxidase (HRP) and a thionine (Th)-coadsorbed CF (HRP/Th-CF)-based bioelectrocatalytic flow-detector at -0.05V vs. Ag/AgCl. Various adsorption conditions of the UOx (i.e., pH of the adsorption solution, type and concentration of the buffer used as the adsorption solvent, UOx concentration and adsorption time) and the operational conditions of the UOx-CF and HRP/Th-CF-coupled flow-biosensor (i.e., carrier flow rate and carrier pH) were optimized to obtain highly sensitive, selective and stable peak current responses to uric acid. The analytical performance of the UOx-CF and HRP/Th-CF-coupled flow biosensor for uric acid was as follows: sensitivity, 0.25μA/uM; linear range, 0.3-20μM; lower detection limit, 0.18μM; and sample throughput, ca. 30-90 samples/h. The resulting amperometric flow-biosensor for uric acid allowed the determination of uric acid in highly diluted body fluids (human serum and urine), and the analytical results obtained by the present biosensor were in fairly good agreement with those obtained by conventional enzyme-based spectrophotometry.
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ABSTRACT: The development of a noninvasive test for uric acid has been the holy grail of uric acid detection research over the last decade. In the present work, a novel matrix comprising of a NiO thin film (a biocompatible material) loaded with Ni microdiscs was prepared on an ITO-coated glass substrate (Ni/NiO/ITO) with the help of RF sputtering for the reagentless detection of uric acid. The bioelectrode was fabricated by immobilizing uricase using a physical adsorption technique on the surface of the Ni/NiO/ITO electrode. The prepared matrix was found to be efficient in sensing biological processes occurring on the surface of the bioelectrode (Ur/Ni/NiO/ITO) in the presence of the analyte (uric acid) to obtain an electronic output. The biosensor exhibits a high sensitivity (431.09 μA mM(-1)), low Km value (0.15 mM), high apparent enzyme activity (5.07 × 10(-2) units per cm(2)), high shelf life (20 weeks) and good selectivity for the detection of uric acid over a wide concentration range (0.05 mM to 1 mM) without any external mediator in the PBS buffer. The obtained results are encouraging for the realization of a reagentless uric acid biosensor with efficient sensing response characteristics.The Analyst 07/2014; · 3.91 Impact Factor
- Materials 01/2014; 7(2). · 1.88 Impact Factor
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ABSTRACT: Articles related to the adsorption of proteins to nanomaterials are reviewed.•Examples from the last 10 years are discussed focused on biosensors.•Article includes general information about the proteins and emerging trends.Analytica Chimica Acta 10/2014; · 4.52 Impact Factor