title: Functionality of dielectrophoretically immobilized enzyme molecules.
abstract: The enzyme horseradish peroxidase has been immobilized on nanoelectrode arrays by alternating current dielectrophoresis (DEP). Preservation of its enzymatic function after field application was demonstrated by oxidizing dihydrorhodamine 123 with hydrogen peroxide as co-oxidant to create its fluorescent form, rhodamine 123 (Rh123). Localization of the fluorescently labeled enzyme and its product was conducted by fluorescence microscopy. Nanoelectrodes were prepared as tungsten pins arranged in square arrays. Experimental parameters for dielectrophoretic immobilization were optimized for even enzyme distribution and for enzymatic efficiency. Enzyme activity was quantified by determination of fluorescence intensities of immobilized enzyme molecules and of Rh123 produced. These results demonstrate that DEP can be applied to immobilize enzyme molecules while retaining their activity and rendering any chemical modifications unnecessary. This introduces a novel way for the preparation of bioactive surfaces for processes such as biosensing.
authors.affiliation: Fraunhofer Institute for Biomedical Engineering (IBMT), Branch Potsdam-Golm, Potsdam, Germany.
authors.forename: Eva-Maria
authors.initials: EM
authors.lastname: Laux
authors.forename: Udo C
authors.initials: UC
authors.lastname: Kaletta
authors.forename: Frank F
authors.initials: FF
authors.lastname: Bier
authors.forename: Christian
authors.initials: C
authors.lastname: Wenger
authors.forename: Ralph
authors.initials: R
authors.lastname: Hölzel
mesh_terms: D004586:Electrophoresis / Q000379:methods*; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000737:chemistry; D008839:Microelectrodes; D036103:Nanotechnology / Q000295:instrumentation*; D020112:Rhodamine 123 / Q000737:chemistry
chemical_list: D004800:Enzymes, Immobilized; D005456:Fluorescent Dyes; D020112:Rhodamine 123
keywords: Catalytic activity; Dielectrophoresis; Enzymes; Immobilization; Nanoelectrodes