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Catecholamine detection using enzymatic amplification
Authors:F. Lisdat,U. Wollenberger,A. Makower,H. Hö  rtnagl,D. Pfeiffer,F.W. Scheller
Affiliation:

a Institute of Biochemistry and Molecular Physiology, University of Potsdam, c/o Max-Delbrück-Centrum, Robert-Rössle-Str. 10, 13122, Berlin, Germany

b Institute of Pharmacology and Toxicology, Charitè, Humboldt-University Berlin Dorotheenstr. 94 10098, Berlin, Germany

c BST Bio Sensor Technology Berlin, Buchholzer Str. 55-61, 13156, Berlin, Germany

Abstract:Different amplification sensors based on the substrate recycling principle were investigated with respect to their applicability to catecholamine detection. In the bioelectrocatalytic approach, glassy carbon electrodes were modified by laccase or a PQQ-dependent glucose dehydrogenase. Substrate recycling occurs and the detection limit is in the lower nanomolar concentration range (e.g. 10 nM dopamine and 1 nM noradrenaline for the laccase- and glucose dehydrogenase-modified electrodes, respectively). Combinations of glucose dehydrogenase with laccase or tyrosinase were investigated as bienzymatic probes. Among the systems we studied, the laccase/glucose dehydrogenase sensor is the most sensitive (detection limit: 0·5 nM adrenaline). The selectivities of the different sensor systems are discussed. Application of the laccase/glucose dehydrogenase electrode in different media (i.e. brain homogenate, heart effluate) was successfully shown. For samples with high concentrations of interfering substances (uric and ascorbic acid), the interferences can be effectively removed using enzymatic methods.
Keywords:catecholamines   biosensor   substrate recycling   laccase   tyrosinase   glucose dehydrogenase   carbon electrode
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