A new approach for the development of electrochemical biosensors using tobacco mosaic virus (TMV) nanotubes and coat protein (CP) aggregates as enzyme nanocarriers is presented. The generic approach has been demonstrated by realizing amperometric glucose sensors combining an array of Pt electrodes loaded with glucose oxidase (GOD)-modified TMV nanotubes and CPs as a model system. The presence of TMV nanotubes or CPs on the sensor surface allows binding of a high amount of enzymes without substantial loss of their activity. The obtained results underline the potential of integrating biomolecule/virus hybrids with electronic transducers for applications in biosensorics.

An amperometric enzyme biosensor has been applied for the detection of adrenaline. The adrenaline biosensor has been prepared by modification of an oxygen electrode with the enzyme laccase that operates at a broad pH range between pH 3.5 to pH 8. The enzyme molecules were immobilized via cross- linking with glutaraldehyde. The sensitivity of the developed adrenaline biosensor in different pH buffer solutions has been studied. (C) 2015 The Authors. Published by Elsevier Ltd.

A sensor system for investigating (bio)degradationprocesses of polymers is presented. The system utilizes semiconductor field-effect sensors and is capable of monitoring the degradation process in-situ and in real-time. The degradation of the polymer poly(d,l-lactic acid) is exemplarily monitored in solutions with different pH value, pH-buffer solution containing the model enzyme lipase from Rhizomucormiehei and cell-culture medium containing supernatants from stimulated and non-stimulated THP-1-derived macrophages mimicking activation of the immune system.

A semiconductor field-effect device has been used for an enzymatically catalyzed degradation of biopolymers for the first time. This novel technique is capable to monitor the degradation process of multiple samples in situ and in real-time. As model system, the degradation of the biopolymer poly(D, L-lactic acid) has been monitored in the degradation medium containing the enzyme lipase from Rhizomucor miehei. The obtained results demonstrate the potential of capacitive field-effect sensors for degradation studies of biodegradable polymers.

The characterization of the degradation kinetics of biodegradable polymers is mandatory with regard to their proper application. In the present work, polymer-modified electrolyte–insulator–semiconductor (PMEIS) field-effect sensors have been applied for in-situ monitoring of the pH-dependent degradation kinetics of the commercially available biopolymer poly(d,l-lactic acid) (PDLLA) in buffer solutions from pH 3 to pH 13. PDLLA films of 500nm thickness were deposited on the surface of an Al–p-Si–SiO2–Ta2O5 structure from a polymer solution by means of spin-coating method. The PMEIS sensor is, in principle, capable to detect any changes in bulk, surface and interface properties of the polymer induced by degradation processes. A faster degradation has been observed for PDLLA films exposed to alkaline solutions (pH 9, pH 11 and pH 13).

High-k perovskite oxide of barium strontium titanate (BST) represents a very attractive multi-functional transducer material for the development of (bio-)chemical sensors for liquids. In this work, BST films have been applied as a sensitive transducer material for a label-free detection of adsorbed charged macromolecules (positively charged polyelectrolytes) and concentration of hydrogen peroxide vapor as well as protection insulator layer for a contactless electrolyte-conductivity sensor. The experimental results of characterization of individual sensors are presented. Special emphasis is devoted towards the development of a capacitively-coupled contactless electrolyte-conductivity sensor.

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.

A miniaturized capacitively coupled contactless conductivity detection ((CD)-D-4) sensor based on high-k perovskite oxide of barium strontium titanate (BST) has been implemented for the first time. The BST films(similar to 120 nm thick) of Ba0.25Sr0.75TiO3 composition were prepared on a p-Si-SiO2-Pt structure by pulsed laser deposition technique using BST targets fabricated by the self-propagating high-temperature synthesis method. The Pt electrodes were buried into the SiO2 layer to obtain a planar structure. For comparison, contact-mode electrolyte-conductivity (EC) sensors without the protective BST layer were also fabricated. To study the influence of the protective BST layer, both sensors were characterized in electrolyte solutions with various conductivities using two-and four-electrode operation modes. The impedance spectra were recorded in a frequency range from 1 Hz to 1 MHz. An equivalent circuit of the (CD)-D-4 sensor is discussed as well.Both, the EC and (CD)-D-4 sensor, demonstrate nearly identical sensor characteristics. The obtained results clearly show the benefits of the use of the BST-based (CD)-D-4 sensor in a four-electrode configuration for contactless conductivity measurements. A linear dependence between the measured conductance and the electrolyte conductivity is obtained in a wide range of electrolyte conductivity from 0.3 mS/cm to 50 mS/cm. Moreover, typical problems associated with contact-mode EC detection such as the effect of possible redox processes, contamination and fouling of electrodes during continuous measurements can be minimized, thus, enhancing the life-time of conductivity sensors considerably. (C) 2014 Elsevier B.V. All rights reserved.

In vitro studies of the degradation kinetic of biopolymers are essential for the design and optimization of implantable biomedical devices. In the presented work, a field-effect capacitive sensor has been applied for the real-time and in situ monitoring of degradation processes of biopolymers for the first time. The polymer-covered field-effect sensor is, in principle, capable to detect any changes in bulk, surface and interface properties of the polymer induced by degradation processes. The feasibility of this approach has been experimentally proven by using the commercially available biomedical polymer poly(D,L-lactic acid) (PDLLA) as a model system. PDLLA films of different thicknesses were deposited on the Ta2O5-gate surface of the field-effect structure from a polymer solution by means of spin-coating method. The polymer-modified field-effect sensors have been characterized by means of capacitance-voltage and impedance-spectroscopy method. The degradation of the PDLLA was accelerated by changing the degradation medium from neutral (pH 7.2) to alkaline (pH 9) condition, resulting in drastic changes in the capacitance and impedance spectra of the polymer-modified field-effect sensor. (C) 2012 Elsevier B. V. All rights reserved.

For the development of new biopolymers and implantable biomedical devices with predicted biodegradability, simple, non-destructive, fast and inexpensive techniques capable for real-time in situ testing of the degradation kinetics of polymers are highly appreciated. In this work, a capacitive field-effect electrolyte-insulator-semiconductor (EIS) sensor has been applied for real-time in situ monitoring of degradation of thin poly(D,L-lactic acid) (PDLLA) films over a long-time period of one month. Generally, the polymer-modified EIS (PMEIS) sensor is capable of detecting any changes in the bulk, surface and interface properties of the polymer (e.g., thickness, coverage, dielectric constant, surface potential) induced by degradation processes. The time-dependent capacitance-voltage (C-V) characteristics of PMEIS structures were used as an indicator of the polymer degradation. To accelerate the PDLLA degradation, experiments were performed in alkaline buffer solution of pH 10.6. The results of these degradation measurements with the EIS sensor were verified by the detection of lactic acid (product of the PDLLA degradation) in the degradation medium. In addition, the micro-structural and morphological changes of the polymer surface induced by the polymer degradation have been systematically studied by means of scanning-electron microscopy, atomic-force microscopy, optical microscopy, and contact-angle measurements. (C) 2013 Elsevier Ltd. All rights reserved.

Two types of microvalves based on temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm) and pH-responsive poly(sodium acrylate) (PSA) hydrogel films have been developed and tested. The PNIPAAm and PSA hydrogel films were prepared by means of in situ photopolymerization directly inside the fluidic channel of a microfluidic chip fabricated by combining Si and SU-8 technologies. The swelling/shrinking properties and height changes of the PNIPAAm and PSA films inside the fluidic channel were studied at temperatures of deionized water from 14 to 36 degrees C and different pH values (pH 3-12) of Titrisol buffer, respectively. Additionally, in separate experiments, the lower critical solution temperature (LCST) of the PNIPAAm hydrogel was investigated by means of a differential scanning calorimetry (DSC) and a surface plasmon resonance (SPR) method. Mass-flow measurements have shown the feasibility of the prepared hydrogel films to work as an on-chip integrated temperature- or pH-responsive microvalve capable to switch the flow channel on/off. (c) 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

A microfluidic chip integrating amperometric enzyme sensors for the detection of glucose, glutamate and glutamine in cell-culture fermentation processes has been developed. The enzymes glucose oxidase, glutamate oxidase and glutaminase were immobilized by means of cross-linking with glutaraldehyde on platinum thin-film electrodes integrated within a microfluidic channel. The biosensor chip was coupled to a flow-injection analysis system for electrochemical characterization of the sensors. The sensors have been characterized in terms of sensitivity, linear working range and detection limit. The sensitivity evaluated from the respective peak areas was 1.47, 3.68 and 0.28 μAs/mM for the glucose, glutamate and glutamine sensor, respectively. The calibration curves were linear up to a concentration of 20 mM glucose and glutamine and up to 10 mM for glutamate. The lower detection limit amounted to be 0.05 mM for the glucose and glutamate sensor, respectively, and 0.1 mM for the glutamine sensor. Experiments in cell-culture medium have demonstrated a good correlation between the glutamate, glutamine and glucose concentrations measured with the chip-based biosensors in a differential-mode and the commercially available instrumentation. The obtained results demonstrate the feasibility of the realized microfluidic biosensor chip for monitoring of bioprocesses.

Field-effect capacitive electrolyte-insulator-semiconductor (EIS) sensors functionalised with citrate-capped gold nanoparticles (AuNP) have been used for the electrostatic detection of macromolecules by their intrinsic molecular charge. The EIS sensor detects the charge changes in the AuNP/macromolecule hybrids induced by the adsorption or binding events. A feasibility of the proposed detection scheme has been exemplary demonstrated by realising EIS sensors for the detection of poly-D-lysine molecules.

A silicon-based biosensor chip consisting of an array of amperometric enzyme sensors has been developed for monitoring the concentration of glutamate and glutamine in cell-culture fermentation processes. The glutamate sensor was constructed by immobilising glutamate oxidase, while the glutamine sensor by sequential coupling of glutaminase and glutamate oxidase. The enzymes were immobilised by means of cross-linking with glutaraldehyde on the surface of the patterned platinum thin-film electrodes. The developed biosensors have been electrochemically characterised in solutions with different concentrations of glutamate and glutamine in terms of sensitivity, response time, linear working range and lifetime. A high sensitivity of 96nA/mM and 100nA/mM was registered for the glutamate and glutamine sensors, respectively. The preliminary experiments in cell-culture medium have shown a good correlation between the glutamine and glutamate concentrations measured with the biosensor chip and the commercially available biochemistry analyser. The obtained results demonstrate the feasibility of the realised biosensor chip for monitoring the glutamine and glutamate concentrations in fermentation processes.

This paper describes the design and characterization of a silicon based sensor chip for monitoring of fermentation processes. The sensor chip consists of three sensors using different transducer principles. A capacitive electrolyte-insulator-semiconductor (EIS) field-effect structure with Ta2O5 as gate material was utilized as pH sensor. An electrolyte conductivity sensor was realized by measuring the impedance between two interdigitated electrodes (IDE). A platinum thermistor was included for temperature measurements. The EIS sensor was integrated into a bioreactor and successfully used for an inline pH measurement. The layout of the IDE has been optimized with respect to a high cell constant and a wide detectable conductivity range. The integrated platinum thermistor allowed for temperature compensation of the electrolyte conductivity measurement. (C) 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

In this study, a concept for a silicon-based modular solid-state sensor system for inline multi-parameter monitoring of cell-culture fermentation processes is presented. The envisaged multi-parameter sensor system consists of two identical sensor modules and is intended for continuous quantification of up to five (bio-)chemical and physical parameters, namely, glucose and glutamine concentration, pH value, electrolyte conductivity and temperature by applying different transducer principles and/or different operation modes. Experimental results for the field-effect electrolyte-insulator-semiconductor (EIS) sterilisable pH sensor and electrolyte conductivity sensor based on interdigitated electrodes are presented. The ongoing autoclaving does not have any significant impact on the pH-sensitive properties of a Ta2O5-gate EIS sensor. Even after 30 autoclaving cycles, the pH sensors show a clear pH response and nearly linear calibration curve with a slope of 57±1mV/pH. Additional scanning electron microscopy and ellipsometric investigations do not show any visible surface degradation or changes in the thickness of the pH-sensitive Ta2O5 layer. The preliminary results demonstrate the suitability of the developed EIS sensor for an inline pH measurement during a fermentation process. In addition, interdigitated electrodes of different geometries serving as electrolyte conductivity sensor have been tested for measurements in relatively high ionic-strength solutions.

A capacitive field-effect EDIS (electrolyte–diamond–insulator–semiconductor) sensor with improved pH and penicillin sensitivity has been realised using a nanocrystalline-diamond (NCD) film as sensitive gate material. The NCD growth process on SiO2 as well as an additional surface treatment in oxidising medium have been optimised to provide high pH-sensitive, non-porous O-terminated films without damage of the underlying SiO2 layer. The surface morphology of O-terminated NCD thin films and the layer structure of EDIS sensors have been studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) methods. To establish the relative coverage of the surface functional groups generated by the oxidation of NCD surfaces, X-ray photoelectron spectroscopy analysis was carried out. The hydrophilicity of NCD thin films has been studied by water contact-angle measurements. A nearly Nernstian pH sensitivity of 54–57mV/pH has been observed for O-terminated NCD films treated in an oxidising boiling mixture for 80min and in oxygen plasma. The high pH-sensitive properties of O-terminated NCD have been used to develop an EDIS-based penicillin biosensor. A freshly prepared penicillin biosensor possesses a high sensitivity of 85mV/decade in the concentration range of 0.1–2.5mM penicillin G. The lower detection limit is 5μM.