TY - JOUR A1 - Ribitsch, D. A1 - Heumann, S. A1 - Karl, W. A1 - Gerlach, J. A1 - Leber, R. A1 - Birner-Gruenberger, R. A1 - Gruber, K. A1 - Eiteljoerg, I. A1 - Remler, P. A1 - Siegert, Petra A1 - Lange, J. A1 - Maurer, Karl-Heinz A1 - Berg, G. A1 - Guebitz, G. M. A1 - Schwab, H. T1 - Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli JF - Journal of biotechnology N2 - A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli. Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3. From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45 °C. Specific activity of StmPr2 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 17 ± 2 U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2. KW - Alginate beads KW - Stenotrophomonas maltophilia KW - Detergent protease Y1 - 2012 U6 - https://doi.org/10.1016/j.jbiotec.2011.09.025 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - 157 IS - 1 SP - 140 EP - 147 PB - Elsevier CY - Amsterdam ER - TY - CHAP A1 - Bohrn, Ulrich A1 - Mucha, Andreas A1 - Werner, Frederik A1 - Stütz, Evamaria A1 - Bäcker, Matthias A1 - Krumbe, Christoph A1 - Schienle, Meinrad A1 - Fleischer, Maximilian A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Detection of toxic chromium species in water using cellbased sensor systems Y1 - 2012 SN - 978-3-9813484-2-2 U6 - https://doi.org/10.5162/IMCS2012/P2.1.14 SP - 1364 EP - 1367 ER - TY - CHAP A1 - Bohrn, Ulrich A1 - Stütz, Evamaria A1 - Fleischer, Maximilian A1 - Schöning, Michael Josef A1 - Wagner, Patrick T1 - Living cell-based gas sensor system for the detection of acetone in air Y1 - 2012 SN - 978-3-9813484-2-2 U6 - https://doi.org/10.5162/IMCS2012/3.2.3 SP - 269 EP - 272 ER - TY - CHAP A1 - Takenaga, Shoko A1 - Werner, Frederik A1 - Sawada, Kazuaki A1 - Schöning, Michael Josef T1 - Comparison of label-free ACh image sensors based on CCD and LAPS Y1 - 2012 SN - 978-3-9813484-2-2 U6 - https://doi.org/10.5162/IMCS2012/4.2.6 SP - 356 EP - 359 ER - TY - JOUR A1 - Henken, F. E. A1 - Oosterhuis, K. A1 - Öhlschläger, Peter A1 - Bosch, L. A1 - Hooijberg, E. A1 - Haanen, J. B. A. G. A1 - Steenbergen, R. D. M. T1 - Preclinical safety evaluation of DNA vaccines encoding modified HPV16 E6 and E7 JF - Vaccine N2 - Persistent infection with high-risk human papillomaviruses (hrHPV) can result in the formation of anogenital cancers. As hrHPV proteins E6 and E7 are required for cancer initiation and maintenance, they are ideal targets for immunotherapeutic interventions. Previously, we have described the development of DNA vaccines for the induction of HPV16 E6 and E7 specific T cell immunity. These vaccines consist of ‘gene-shuffled’ (SH) versions of HPV16 E6 and E7 that were fused to Tetanus Toxin Fragment C domain 1 (TTFC) and were named TTFC-E6SH and TTFC-E7SH. Gene-shuffling was performed to avoid the risk of inducing malignant transformation at the vaccination site. Here, we describe the preclinical safety evaluation of these candidate vaccines by analysis of their transforming capacity in vitro using established murine fibroblasts (NIH 3T3 cells) and primary human foreskin keratinocytes (HFKs). We demonstrate that neither ectopic expression of TTFC-E6SH and TTFC-E7SH alone or in combination enabled NIH 3T3 cells to form colonies in soft agar. In contrast, expression of HPV16 E6WT and E7WT alone or in combination resulted in effective transformation. Similarly, retroviral transduction of HFKs from three independent donors with both TTFC-E6SH and TTFC-E7SH alone or in combination did not show any signs of immortalization. In contrast, the combined expression of E6WT and E7WT induced immortalization in HFKs from all donors. Based on these results we consider it justified to proceed to clinical evaluation of DNA vaccines encoding TTFC-E6SH and TTFC-E7SH in patients with HPV16 associated (pre)malignancies. Y1 - 2012 U6 - https://doi.org/10.1016/j.vaccine.2012.04.013 SN - 0264-410X VL - 30 IS - 28 SP - 4259 EP - 4266 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Immel, Timo A1 - Grützke, Martin A1 - Späte, Anne-Katrin A1 - Groth, Ulrich A1 - Öhlschläger, Peter A1 - Huhn, Thomas T1 - Synthesis and X-ray structure analysis of a heptacoordinate titanium(IV)-bis-chelate with enhanced in vivo antitumor efficacy JF - Chemical Communications N2 - Chelate stabilization of a titanium(IV)–salan alkoxide by ligand exchange with 2,6-pyridinedicarboxylic acid (dipic) resulted in heptacoordinate complex 3 which is not redox-active, stable on silica gel and has increased aqueous stability. 3 is highly toxic in HeLa S3 and Hep G2 and has enhanced antitumor efficacy in a mouse cervical-cancer model. Y1 - 2012 U6 - https://doi.org/10.1039/C2CC31624B SN - 1364-548X VL - 48 IS - 46 SP - 5790 EP - 5792 PB - Royal Society of Chemistry CY - Cambridge ER - TY - JOUR A1 - Borgmeier, Claudia A1 - Bongaerts, Johannes A1 - Meinhardt, Friedhelm T1 - Genetic analysis of the Bacillus licheniformis degSU operon and the impact of regulatory mutations on protease production JF - Journal of biotechnology N2 - Disruption experiments targeted at the Bacillus licheniformis degSU operon and GFP-reporter analysis provided evidence for promoter activity immediately upstream of degU. pMutin mediated concomitant introduction of the degU32 allele – known to cause hypersecretion in Bacillus subtilis – resulted in a marked increase in protease activity. Application of 5-fluorouracil based counterselection through establishment of a phosphoribosyltransferase deficient Δupp strain eventually facilitated the marker-free introduction of degU32 leading to further protease enhancement achieving levels as for hypersecreting wild strains in which degU was overexpressed. Surprisingly, deletion of rapG – known to interfere with DegU DNA-binding in B. subtilis – did not enhance protease production neither in the wild type nor in the degU32 strain. The combination of degU32 and Δupp counterselection in the type strain is not only equally effective as in hypersecreting wild strains with respect to protease production but furthermore facilitates genetic strain improvement aiming at biological containment and effectiveness of biotechnological processes. KW - Marker-free mutagenesis KW - Extracellular enzymes KW - Uracil-phosphoribosyltransferase KW - Hypersecretion Y1 - 2012 U6 - https://doi.org/10.1016/j.jbiotec.2012.02.011 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - 159 IS - 1-2 SP - 12 EP - 20 PB - Elsevier CY - Amsterdam ER - TY - CHAP A1 - Weil, M. A1 - Poghossian, Arshak A1 - Schöning, Michael Josef A1 - Cherstvy, A. T1 - Electrical monitoring of layer-by-layer adsorption of oppositely charged macromolecules by means of capacitive field-effect devices Y1 - 2012 SN - 978-3-9813484-2-2 U6 - https://doi.org/10.5162/IMCS2012/P2.5.2 SP - 1575 EP - 1578 ER - TY - JOUR A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Cherstvy, Andrey G. A1 - Pedraza, Angela M. A1 - Ingebrandt, Sven A1 - Schöning, Michael Josef T1 - Label-free electrical detection of DNA by means of field-effect nanoplate capacitors: Experiments and modeling JF - Physica Status Solidi (a) N2 - Label-free electrical detection of consecutive deoxyribonucleic acid (DNA) hybridization/denaturation by means of an array of individually addressable field-effect-based nanoplate silicon-on-insulator (SOI) capacitors modified with gold nanoparticles (Au-NP) is investigated. The proposed device detects charge changes on Au-NP/DNA hybrids induced by the hybridization or denaturation event. DNA hybridization was performed in a high ionic-strength solution to provide a high hybridization efficiency. On the other hand, to reduce the screening of the DNA charge by counter ions and to achieve a high sensitivity, the sensor signal induced by the hybridization and denaturation events was measured in a low ionic-strength solution. High sensor signals of about 120, 90, and 80 mV were registered after the DNA hybridization, denaturation, and re-hybridization events, respectively. Fluorescence microscopy has been applied as reference method to verify the DNA immobilization, hybridization, and denaturation processes. An electrostatic charge-plane model for potential changes at the gate surface of a nanoplate field-effect sensor induced by the DNA hybridization has been developed taking into account both the Debye length and the distance of the DNA charge from the gate surface. Y1 - 2012 U6 - https://doi.org/10.1002/pssa.201100710 SN - 1862-6319 VL - 209 SP - 925 EP - 934 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Huck, Christina A1 - Poghossian, Arshak A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Combined amperometric/field-effect sensor for the detection of dissolved hydrogen JF - Sensors and actuators B: Chemical N2 - Real-time and reliable monitoring of the biogas process is crucial for a stable and efficient operation of biogas production in order to avoid digester breakdowns. The concentration of dissolved hydrogen (H₂) represents one of the key parameters for biogas process control. In this work, a one-chip integrated combined amperometric/field-effect sensor for monitoring the dissolved H₂ concentration has been developed for biogas applications. The combination of two different transducer principles might allow a more accurate and reliable measurement of dissolved H₂ as an early warning indicator of digester failures. The feasibility of the approach has been demonstrated by simultaneous amperometric/field-effect measurements of dissolved H₂ concentrations in electrolyte solutions. Both, the amperometric and the field-effect transducer show a linear response behaviour in the H₂ concentration range from 0.1 to 3% (v/v) with a slope of 198.4 ± 13.7 nA/% (v/v) and 14.9 ± 0.5 mV/% (v/v), respectively. Y1 - 2012 U6 - https://doi.org/10.1016/j.snb.2012.10.050 SN - 0925-4005 N1 - Part of special issue "Selected Papers from the 14th International Meeting on Chemical Sensors" VL - 187 SP - 168 EP - 173 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Poghossian, Arshak A1 - Weil, M. H. A1 - Bäcker, Matthias A1 - Mayer, D. A1 - Schöning, Michael Josef T1 - Field-effect Devices Functionalised with Gold-Nanoparticle/Macromolecule Hybrids: New Opportunities for a Label-Free Biosensing JF - Procedia Engineering N2 - 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. Y1 - 2012 U6 - https://doi.org/10.1016/j.proeng.2012.09.136 SN - 1877-7058 N1 - Part of special issue "26th European Conference on Solid-State Transducers, EUROSENSOR 2012" IS - 47 SP - 273 EP - 276 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Schusser, Sebastian A1 - Poghossian, Arshak A1 - Bäcker, Matthias A1 - Leinhos, Marcel A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Characterization of biodegradable polymers with capacitive field-effect sensors JF - Sensors and actuators B: Chemical N2 - 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. KW - Impedance spectroscopy KW - C–V method KW - Real-time monitoring KW - Poly(d,l-lacticacid) KW - (Bio)degradation KW - Field-effect sensor Y1 - 2012 U6 - https://doi.org/10.1016/j.snb.2012.07.099 SN - 0925-4005 N1 - Part of special issue "Selected Papers from the 14th International Meeting on Chemical Sensors" VL - 187 SP - 2 EP - 7 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bäcker, Matthias A1 - Raue, Markus A1 - Schusser, Sebastian A1 - Jeitner, C. A1 - Breuer, L. A1 - Wagner, P. A1 - Poghossian, Arshak A1 - Förster, Arnold A1 - Mang, Thomas A1 - Schöning, Michael Josef T1 - Microfluidic chip with integrated microvalves based on temperature- and pH-responsive hydrogel thin films JF - Physica Status Solidi (a) N2 - 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 °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. Y1 - 2012 U6 - https://doi.org/10.1002/pssa.201100763 SN - 1862-6319 VL - 209 IS - 5 SP - 839 EP - 845 PB - Wiley-VCH CY - Weinheim ER - TY - JOUR A1 - Bohrn, Ulrich A1 - Stütz, Evamaria A1 - Fleischer, Maximilian A1 - Schöning, Michael Josef A1 - Wagner, Patrick T1 - Using a cell-based gas biosensor for investigation of adverse effects of acetone vapors in vitro JF - Biosensors and Bioelectronics. 40 (2013), H. 1 Y1 - 2013 SN - 0956-5663 SP - 393 EP - 400 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Hennemann, Jörg A1 - Kohl, Claus-Dieter A1 - Reisert, Steffen A1 - Kirchner, Patrick A1 - Schöning, Michael Josef T1 - Copper oxide nanofibres for detection of hydrogen peroxide vapour at high concentrations JF - physica status solidi (a) N2 - We present a sensor concept based on copper(II)oxide (CuO) nanofibres for the detection of hydrogen peroxide (H2O2) vapour in the percent per volume (% v/v) range. The fibres were produced by using the electrospinning technique. To avoid water condensation in the pores, the fibres were initially modified by an exposure to H2S to get an enclosed surface. By a thermal treatment at 350 °C the fibres were oxidised back to CuO. Thereby, the visible pores disappear which was verified by SEM analysis. The fibres show a decrease of resistance with increasing H2O2 concentration which is due to the fact that hydrogen peroxide is an oxidising gas and CuO a p-type semiconductor. The sensor shows a change of resistance within the minute range to the exposure until the maximum concentration of 6.9% v/v H2O2. At operating temperatures below 450 °C the corresponding sensor response to a concentration of 4.1% v/v increases. The sensor shows a good reproducibility of the signal at different measurements. CuO seems to be a suitable candidate for the detection of H2O2 vapour at high concentrations. Resistance behaviour of the sensor under exposure to H2O2 vapours between 2.3 and 6.9% v/v at an operating temperature of 450 °C. Y1 - 2013 U6 - https://doi.org/10.1002/pssa.201200775 SN - 1862-6319 VL - 210 IS - 5 SP - 859 EP - 863 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Reisert, Steffen A1 - Schneider, Benno A1 - Geissler, Hanno A1 - Gompel, Matthias van A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Multi-sensor chip for the investigation of different types of metal oxides for the detection of H2O2 in the ppm range JF - physica status solidi (a) N2 - In this work, a multi-sensor chip for the investigation of the sensing properties of different types of metal oxides towards hydrogen peroxide in the ppm range is presented. The fabrication process and physical characterization of the multi-sensor chip are described. Pure SnO2 and WO3 as well as Pd- and Pt-doped SnO2 films are characterized in terms of their sensitivity to H2O2. The sensing films have been prepared by drop-coating of water-dispensed nano-powders. A physical characterization, including scanning electron microscopy and X-ray diffraction analysis of the deposited metal-oxide films, was done. From the measurements in hydrogen peroxide atmosphere, it could be shown, that all of the tested metal oxide films are suitable for the detection of H2O2 in the ppm range. The highest sensitivity and reproducibility was achieved using Pt-doped SnO2. Calibration plot of a SnO2, WO3, Pt-, and Pd-doped SnO2 gas sensor for H2O2 concentrations in the ppm range. Y1 - 2013 SN - 1862-6319 VL - 210 IS - 5 SP - 898 EP - 904 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Kirchner, Patrick A1 - Oberländer, Jan A1 - Suso, Henri-Pierre A1 - Rysstad, Gunnar A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Towards a wireless sensor system for real-time H2O2 monitoring in aseptic food processes JF - Physica status solidi (a) N2 - A wireless sensor system based on the industrial ZigBee standard for low-rate wireless networking was developed that enables real-time monitoring of gaseous H2O2 during the package sterilization in aseptic food processes. The sensor system consists of a remote unit connected to a calorimetric gas sensor, which was already established in former works, and an external base unit connected to a laptop computer. The remote unit was built up by an XBee radio frequency (RF) module for data communication and a programmable system-on-chip controller to read out the sensor signal and process the sensor data, whereas the base unit is a second XBee RF module. For the rapid H2O2 detection on various locations inside the package that has to be sterilized, a novel read-out strategy of the calorimetric gas sensor was established, wherein the sensor response is measured within the short sterilization time and correlated with the present H2O2 concentration. In an exemplary measurement application in an aseptic filling machinery, the suitability of the new, wireless sensor system was demonstrated, wherein the influence of the gas velocity on the H2O2 distribution inside a package was determined and verified with microbiological tests. KW - calorimetric gas sensor;hydrogen peroxide;wireless sensor system Y1 - 2013 U6 - https://doi.org/10.1002/pssa.201200920 SN - 1862-6319 VL - 210 IS - 5 SP - 877 EP - 883 PB - Wiley CY - Weinheim ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Ichimura, Hiroki A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Chemical imaging of the concentration profile of ion diffusion in a microfluidic channel JF - Sensors and actuators. B: Chemical N2 - The chemical imaging sensor is a device to visualize the spatial distribution of chemical species based on the principle of LAPS (light-addressable potentiometric sensor), which is a field-effect chemical sensor based on semiconductor. In this study, the chemical imaging sensor has been applied to investigate the ion profile of laminar flows in a microfluidic channel. The chemical images (pH maps) were collected in a Y-shaped microfluidic channel while injecting HCl and NaCl solutions into two branches. From the chemical images, it was clearly observed that the injected solutions formed laminar flows in the channel. In addition, ion diffusion across the laminar flows was observed, and the diffusion coefficient could be derived by fitting the pH profiles to the Fick's equation. Y1 - 2013 U6 - https://doi.org/10.1016/j.snb.2013.04.057 SN - 1873-3077 (E-Journal); 0925-4005 (Print) N1 - Part of special issue "Selected Papers from the 26th European Conference on Solid-State Transducers" VL - 189 SP - 240 EP - 245 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bohrn, Ulrich A1 - Mucha, Andreas A1 - Werner, Frederik A1 - Trattner, Barbara A1 - Bäcker, Matthias A1 - Krumbe, Christoph A1 - Schienle, Meinrad A1 - Stütz, Evamaria A1 - Schmitt-Landsiedel, Doris A1 - Fleischer, Maximilian A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - A critical comparison of cell-based sensor systems for the detection of Cr (VI) in aquatic environment JF - Sensors and actuators. B: Chemical Y1 - 2013 SN - 1873-3077 (E-Journal); 0925-4005 (Print) VL - Vol. 182 SP - 58 EP - 65 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Werner, Frederik A1 - Takenaga, Shoko A1 - Taki, Hidenori A1 - Sawada, Kazuaki A1 - Schöning, Michael Josef T1 - Comparison of label-free ACh-imaging sensors based on CCD and LAPS JF - Sensors and Actuators B: Chemical (2012) N2 - Semiconductor-based chemical imaging sensors, like the light-addressable potentiometric sensor (LAPS) or the pH-imaging sensor based on a charge-coupled device (CCD), are becoming a powerful tool for label-free imaging of biological phenomena. We have proposed a polyion-based enzymatic membrane to develop an acetylcholine (ACh) imaging sensor for neural cell-activity observations. In this study, a CCD-type ACh-imaging sensor and a LAPS-type ACh-imaging sensor were fabricated and the prospect of both sensors was clarified by making a comparison of their basic characteristics. Y1 - 2013 SN - 0925-4005 VL - 177 SP - 745 EP - 752 PB - Elsevier CY - Amsterdam ER -