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A new approach for a label-free electrical detection of DNA hybridization and denaturation using an array of individually addressable field-effect nanoplate SOI (silicon-on-insulator) capacitors functionalized with gold nanoparticles is presented. By using a constant-capacitance measuring setup in a differential mode, signal changes of ∼110 mV and ∼70 mV have been registered after the DNA hybridization and denaturation events, respectively.
A novel strategy for enhanced field-effect biosensing using capacitive electrolyte–insulator–semiconductor (EIS) structures functionalised with pH-responsive weak polyelectrolyte/enzyme or dendrimer/enzyme multilayers is presented. The feasibility of the proposed approach is exemplarily demonstrated by realising a penicillin biosensor based on a capacitive p-Si–SiO2 EIS structure functionalised with a poly(allylamine hydrochloride) (PAH)/penicillinase and a poly(amidoamine) dendrimer/penicillinase multilayer. The developed sensors response to changes in both the local pH value near the gate surface and the charge of macromolecules induced via enzymatic reaction, resulting in a higher sensitivity. For comparison, an EIS penicillin biosensor with adsorptively immobilised penicillinase has been also studied. The highest penicillin sensitivity of 100 mV/dec has been observed for the EIS sensor functionalised with the PAH/penicillinase multilayer. The lower and upper detection limit was around 20 µM and 10 mM, respectively. In addition, an incorporation of enzymes in a multilayer prepared by layer-by-layer technique provides a larger amount of immobilised enzymes per sensor area, reduces enzyme leaching effects and thus, enhances the biosensor lifetime (the loss of penicillin sensitivity after 2 months was 10–12%).
Urinary stone formation has been evolved to a widespread disease during the last years. The reason for the formation of urinary stones are little crystals, mostly composed of calcium oxalate, which are formed in human kidneys. The early diagnosis of the risk for urinary stone formation of patients can be determined by the “Bonn-Risk-Index” method based on the potentiometric detection of the Ca2+-ion concentration and an optical determination of the triggered crystallisation of calcium oxalate in unprocessed urine. In this work, miniaturised capacitive field-effect EMIS (electrolyte-membrane-insulator-semiconductor) sensors have been developed for the determination of the Ca2+-ion concentration in human native urine. The Ca2+-sensitive EMIS sensors have been systematically characterised by impedance spectroscopy, capacitance–voltage and constant–capacitance method in terms of sensitivity, signal stability and response time in both CaCl2 solutions and in native urine. The obtained results demonstrate the suitability of EMIS sensors for the measurement of the Ca2+-ion concentration in native urine of patients.
In this contribution, we focus on the detection of toxic gases with living eukaryotic cells. A cell-based gas sensor system, able to measure the effects of direct exposure of gases to cells in real-time, was set up. Impedance data as well as oxygen consumption of Chinese hamster lung fibroblast cells (V79) were analysed upon exposure to carbon monoxide (CO). The CO (diluted in wet synthetic air) affects the cell respiration as indicated by an attenuated respiration signal after the CO exposure as well as an instant increase of the capacitive part of the impedance signal during the gas exposure.
pH-sensitive properties of barium strontium titanate (BST) high-k thin films as alternative gate material for field-effect capacitive (bio-)chemical sensors based on an electrolyte-insulator-semiconductor system have been investigated. The BST films of different compositions (Ba0.31Sr0.69TiO3, Ba0.25Sr0.75TiO3 and Mg-doped Ba0.8Sr0.2Mg0.1Ti0.9O3) were deposited by pulsed laser deposition technique from targets fabricated by self-propagating high-temperature synthesis. The realised sensors have been electrochemically characterised by means of impedance-spectroscopy, capacitance–voltage and constant-capacitance method. The sensors possess a Nernstian-like pH sensitivity in the concentration range between pH 3 and 11 with a response time of 5–10 s. An equivalent circuit model for the BST-based capacitive field-effect sensor is discussed.
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.
Bacillus subtilis and Bacillus licheniformis are widely used for the large-scale industrial production of proteins. These strains can efficiently secrete proteins into the culture medium using the general secretion (Sec) pathway. A characteristic feature of all secreted proteins is their N-terminal signal peptides, which are recognized by the secretion machinery. Here, we have studied the production of an industrially important secreted protease, namely, subtilisin BPN′ from Bacillus amyloliquefaciens. One hundred seventy-three signal peptides originating from B. subtilis and 220 signal peptides from the B. licheniformis type strain were fused to this secretion target and expressed in B. subtilis, and the resulting library was analyzed by high-throughput screening for extracellular proteolytic activity. We have identified a number of signal peptides originating from both organisms which produced significantly increased yield of the secreted protease. Interestingly, we observed that levels of extracellular protease were improved not only in B. subtilis, which was used as the screening host, but also in two different B. licheniformis strains. To date, it is impossible to predict which signal peptide will result in better secretion and thus an improved yield of a given extracellular target protein. Our data show that screening a library consisting of homologous and heterologous signal peptides fused to a target protein can identify more-effective signal peptides, resulting in improved protein export not only in the original screening host but also in different production strains.
Realization of a calorimetric gas sensor on polyimide foil for applications in aseptic food industry
(2010)
A calorimetric gas sensor is presented for the monitoring of gas-phase H2O2 at elevated temperature during sterilization processes in aseptic food industry. The sensor consists of two temperature-sensitive thin-film resistances built up on a polyimide foil with a thickness of 25 μm, which are passivated with a layer of SU-8 photo resist and catalytically activated with manganese(IV) oxide. Instead of an active heating structure, the calorimetric sensor utilizes the elevated temperature of an evaporated H2O2 aerosol. In an experimental set-up, the sensor has shown a sensitivity of 4.78 °C/(%v/v) in a H2O2 concentration range of 0 to 10% v/v at an evaporation temperature of 240 ∘C. Furthermore, the sensor possesses the same, unchanged sensor signal even at varied evaporation temperatures of the gas stream. The sensor characterization demonstrates the suitability of the calorimetric gas sensor for monitoring the efficiency of sterilization processes.
In aseptischen Abfüllsystemen wird Wasserstoffperoxid in der Gasphase aufgrund der stark oxidativen Wirkung zur Packstoffentkeimung eingesetzt. Dabei wird die Effizienz der Entkeimung im Wesentlichen von der vorliegenden H2O2-Konzentration im Packstoff bestimmt. Zur Inline-Überwachung der H2O2-Konzentration wurde ein kalorimetrischer Gassensor auf Basis einer flexiblen Polyimidfolie aus temperatursensitiven Dünnschicht-Widerständen und Mangan(IV)-oxid als katalytische Transducerschicht realisiert. Der Sensor weist ein lineares Ansprechverhalten mit einer Sensitivität von 7,15 °C/Vol.-% in einem H2O2-Konzentrationsbereich von 0 bis 8 Vol.-% auf. Weiterhin wurde zur Auslesung des Sensorsignals eine RFID-Elektronik, bestehend aus einem Sensor-Tag und einer Sende-/Empfangseinheit ausgelegt, sowie eine Abfolge des Messzyklus aufgestellt. Im weiteren Verlauf soll der kalorimetrische Gassensor mit der RFID-Elektronik gekoppelt und in eine Testverpackung zur Inline-Überwachung der H2O2-Konzentration in aseptischen Abfüllsystemen implementiert werden.