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In diesem Beitrag werden Ergebnisse der Entwicklung eines modularen festkörperbasierten Sensorsystems für die Überwachung von Zellkulturfermentationen präsentiert. Zur Messung der Elektrolytleitfähigkeit wurde das Layout von Interdigitalelektroden angepasst, um in vergleichsweise gut leitenden Elektrolyten zu messen. Durch Quervernetzung von Glucose-Oxidase mit Glutaraldehyd und Immobilisierung auf einer Platinelektrode wurde ein amperometrischer Glucosesensor mit einem linearen Messbereich von bis zu 2 mM und einer Sensitivität von 168 nA/mM realisiert.
Capacitive field-effect electrolyte-diamond-insulator-semiconductor (EDIS) structures with O-terminated nanocrystalline diamond (NCD) as sensitive gate material have been realized and investigated for the detection of pH, penicillin concentration, and layer-by-layer adsorption of polyelectrolytes. The surface oxidizing procedure of NCD thin films as well as the seeding and NCD growth process on a Si-SiO2 substrate have been improved to provide high pH-sensitive, non-porous thin films without damage of the underlying SiO2 layer and with a high coverage of O-terminated sites. The NCD surface topography, roughness, and coverage of the surface groups have been characterized by SEM, AFM and XPS methods. The EDIS sensors with O-terminated NCD film treated in oxidizing boiling mixture for 45 min show a pH sensitivity of about 50 mV/pH. The pH-sensitive properties of the NCD have been used to develop an EDIS-based penicillin biosensor with high sensitivity (65-70 mV/decade in the concentration range of 0.25-2.5 mM penicillin G) and low detection limit (5 μM). The results of label-free electrical detection of layer-by-layer adsorption of charged polyelectrolytes are presented, too.
Planar and three-dimensional (3D) interdigitated electrodes (IDE) with electrode digits separated by an insulating barrier of different heights were electrochemically characterized and compared in terms of their sensing properties. Due to the impact of the surface resistance, both types of IDE structures display a non-linear behavior in low-ionic strength solutions. The experimental data were fitted to an electrical equivalent circuit and interpreted taking into account the surface-charge-governed properties. The effect of a charged polyelectrolyte layer electrostatically assembled onto the sensor surface on the surface resistance in solutions with different KCl concentration is studied. In case of the same electrode footprint, 3D-IDEs show a larger cell constant and a higher sensitivity to molecular adsorption than that of planar IDEs. The obtained results demonstrate the potential of 3D-IDEs as a new transducer structure for a direct label-free sensing of charged molecules.
The conjunction of (bio-)chemical recognition elements with nanoscale biological building blocks such as virus particles is considered as a very promising strategy for the creation of biohybrids opening novel opportunities for label-free biosensing. This work presents a new approach for the development of biosensors using tobacco mosaic virus (TMV) nanotubes or coat proteins (CPs) as enzyme nanocarriers. Sensor chips combining an array of Pt electrodes loaded with glucose oxidase (GOD)-modified TMV nanotubes or CP aggregates were used for amperometric detection of glucose as a model system for the first time. The presence of TMV nanotubes or CPs on the sensor surface allows binding of a high amount of precisely positioned enzymes without substantial loss of their activity, and may also ensure accessibility of their active centers for analyte molecules. Specific and efficient immobilization of streptavidin-conjugated GOD ([SA]-GOD) complexes on biotinylated TMV nanotubes or CPs was achieved via bioaffinity binding. These layouts were tested in parallel with glucose sensors with adsorptively immobilized [SA]-GOD, as well as [SA]-GOD crosslinked with glutardialdehyde, and came out to exhibit superior sensor performance. The achieved results underline a great potential of an integration of virus/biomolecule hybrids with electronic transducers for future applications in biosensorics and biochips.