TY - JOUR A1 - Wagner, Torsten A1 - Werner, Frederik A1 - Miyamoto, Ko-Ichiro A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - A high-density multi-point LAPS set-up using a VCSEL array and FPGA control JF - Sensors and Actuators B: Chemical. 154 (2011), H. 2 Y1 - 2011 SN - 1873-3077 N1 - EUROSENSORS XXIII SP - 124 EP - 128 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Mimura, Shuhei A1 - Kanoh, Shiníchiro A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Constant-phase-mode operation of the light-addressable potentiometric sensor JF - Sensors and Actuators B: Chemical. 154 (2011), H. 2 Y1 - 2011 SN - 1873-3077 N1 - EUROSENSORS XXIII SP - 119 EP - 123 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Kirchner, Patrick A1 - Li, Bin A1 - Spelthahn, Heiko A1 - Henkel, Hartmut A1 - Schneider, Andreas A1 - Friedrich, Peter A1 - Kolstad, Jens A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Thin-film calorimetric H2O2 gas sensor for the validation of germicidal effectivity in aseptic filling processes JF - Sensors and Actuators B: Chemical. 154 (2011), H. 2 Y1 - 2011 SN - 1873-3077 N1 - EUROSENSORS XXIII SP - 257 EP - 263 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Yoshinobu, Tatsuo A1 - Kanoh, Shin`ichiro A1 - Schöning, Michael Josef T1 - Phase-mode LAPS and its application to chemical imaging JF - Sensors and Actuators B: Chemical. 154 (2011), H. 1 Y1 - 2011 SN - 1873-3077 SP - 28 EP - 32 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bäcker, Matthias A1 - Delle, L. A1 - Poghossian, Arshak A1 - Biselli, Manfred A1 - Zang, Werner A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Electrochemical sensor array for bioprocess monitoring JF - Electrochimica Acta (2011) Y1 - 2011 VL - 56 IS - 26 SP - 9673 EP - 9678 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bäcker, Matthias A1 - Rakowski, D. A1 - Poghossian, Arshak A1 - Biselli, Manfred A1 - Wagner, Patrick A1 - Schöning, Michael Josef T1 - Chip-based amperometric enzyme sensor system for monitoring of bioprocesses by flow-injection analysis JF - Journal of Biotechnology N2 - 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. Y1 - 2013 U6 - https://doi.org/10.1016/j.jbiotec.2012.03.014 SN - 0168-1656 VL - 163 IS - 4 SP - 371 EP - 376 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Spiess, Elmar A1 - Wilfried, Reichardt A1 - Alvarez, Gerardo A1 - Gottrup, Marcus A1 - Öhlschläger, Peter T1 - An Artificial PAP Gene Breaks Self-tolerance and Promotes Tumor Regression in the TRAMP Model for Prostate Carcinoma JF - Molecular Therapy Y1 - 2011 SN - 1525-0016 VL - 20 IS - 3 SP - 555 EP - 564 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 - TY - JOUR A1 - Kirchner, Patrick A1 - Oberländer, Jan A1 - Suco, Henri-Pierre A1 - Rysstad, Gunnar A1 - Schöning, Michael Josef T1 - Monitoring the microbicidal effectiveness of gaseous hydrogen peroxide in sterilisation processes by means of a calorimetric gas sensor JF - Food control N2 - In the present work, a novel method for monitoring sterilisation processes with gaseous H2O2 in combination with heat activation by means of a specially designed calorimetric gas sensor was evaluated. Therefore, the sterilisation process was extensively studied by using test specimens inoculated with Bacillus atrophaeus spores in order to identify the most influencing process factors on its microbicidal effectiveness. Besides the contact time of the test specimens with gaseous H2O2 varied between 0.2 and 0.5 s, the present H2O2 concentration in a range from 0 to 8% v/v (volume percent) had a strong influence on the microbicidal effectiveness, whereas the change of the vaporiser temperature, gas flow and humidity were almost negligible. Furthermore, a calorimetric H2O2 gas sensor was characterised in the sterilisation process with gaseous H2O2 in a wide range of parameter settings, wherein the measurement signal has shown a linear response against the H2O2 concentration with a sensitivity of 4.75 °C/(% v/v). In a final step, a correlation model by matching the measurement signal of the gas sensor with the microbial inactivation kinetics was established that demonstrates its suitability as an efficient method for validating the microbicidal effectiveness of sterilisation processes with gaseous H2O2. KW - hydrogen peroxide KW - sterilisation KW - Bacillus atrophaeus KW - calorimetric gas sensor Y1 - 2012 U6 - https://doi.org/10.1016/j.foodcont.2012.11.048 SN - 0956-7135 VL - 31 IS - 2 SP - 530 EP - 538 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Schusser, Sebastian A1 - Menzel, S. A1 - Bäcker, Matthias A1 - Leinhos, Marcel A1 - Poghossian, Arshak A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Degradation of thin poly(lactic acid) films: characterization by capacitance-voltage, atomic force microscopy, scanning electron microscopy and contact-angle measurements JF - Electrochimica Acta Y1 - 2013 SN - 1873-3859 (E-Journal); 0013-4686 (Print) VL - Vol. 113 SP - 779 EP - 784 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, K. A1 - Kuwabara, Yohei A1 - Kanoh, Shin'ichiro A1 - Yoshinobu, Tatsuo A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Chemical image scanner based on FDM-LAPS JF - Sensors and Actuators B: Chemical. 137 (2009), H. 2 Y1 - 2009 SN - 0925-4005 SP - 533 EP - 538 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Poghossian, Arshak A1 - Ingebrandt, S. A1 - Offenhäusser, A. A1 - Schöning, Michael Josef T1 - Field-effect devices for detecting cellular signals JF - Seminars in Cell & Developmental Biology. 20 (2009), H. 1 Y1 - 2009 SN - 1096-3634 SP - 41 EP - 48 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Pedraza, A. M. A1 - Gandhi, D. A1 - Ingebrandt, S. A1 - Moritz, W. A1 - Schöning, Michael Josef T1 - An array of field-effect nanoplate SOI capacitors for (bio-)chemical sensing JF - Biosensors and Bioelectronics. 26 (2011), H. 6 Y1 - 2011 SN - 0956-5663 SP - 3023 EP - 3028 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Poghossian, Arshak A1 - Malzahn, K. A1 - Abouzar, Maryam H. A1 - Mehndiratta, P. A1 - Katz, E. A1 - Schöning, Michael Josef T1 - Integration of biomolecular logic gates with field-effect transducers JF - Electrochimica Acta. 56 (2011), H. 26 Y1 - 2011 SN - 0013-4686 SP - 9661 EP - 9665 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Abouzar, Maryam H. A1 - Poghossian, Arshak A1 - Razavi, A. A1 - Williams, O. A. A1 - Bijnens, N. A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Characterisation of capacitive field-effect sensors with a nanocrystalline-diamond film as transducer material for multi-parameter sensing JF - Biosensors and Bioelectronics. 24 (2009), H. 5 Y1 - 2009 SN - 0956-5663 N1 - Selected Papers from the Tenth World Congress on Biosensors Shangai, China, May 14-16, 2008 ; Zeitschrift früher u.d.T. : Biosensors SP - 1298 EP - 1304 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Itabashi, Akinori A1 - Kosaka, Naoki A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - High-speed chemical imaging system based on front-side-illuminated LAPS JF - Sensors and actuators B: Chemical N2 - The chemical imaging sensor is a semiconductor-based chemical sensor that can visualize the spatial distribution of specific ions on the sensing surface. The conventional chemical imaging system based on the light-addressable potentiometric sensor (LAPS), however, required a long time to obtain a chemical image, due to the slow mechanical scan of a single light beam. For high-speed imaging, a plurality of light beams modulated at different frequencies can be employed to measure the ion concentrations simultaneously at different locations on the sensor plate by frequency division multiplex (FDM). However, the conventional measurement geometry of back-side illumination limited the bandwidth of the modulation frequency required for FDM measurement, because of the low-pass filtering characteristics of carrier diffusion in the Si substrate. In this study, a high-speed chemical imaging system based on front-side-illuminated LAPS was developed, which achieved high-speed spatiotemporal recording of pH change at a rate of 70 frames per second. Y1 - 2013 U6 - https://doi.org/10.1016/j.snb.2013.03.016 SN - 1873-3077 VL - 182 SP - 315 EP - 321 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Iken, Heiko A1 - Ahlborn, K. A1 - Gerlach, F. A1 - Vonau, W. A1 - Zander, W. A1 - Schubert, J. A1 - Schöning, Michael Josef T1 - Development of redox glasses and subsequent processing by means of pulsed laser deposition for realizing silicon-based thin-film sensors JF - Electrochimica acta Y1 - 2013 SN - 1873-3859 (E-Journal); 0013-4686 (Print) SP - Available online 30.8.2013 PB - Elsevier CY - Amsterdam 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 - JOUR A1 - Wilming, Anja A1 - Begemann, Jens A1 - Kuhne, Stefan A1 - Regestein, Lars A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Maurer, Karl-Heinz A1 - Büchs, Jochen T1 - Metabolic studies of γ-polyglutamic acid production in Bacillus licheniformis by small-scale continuous cultivations JF - Biochemical engineering journal Y1 - 2013 SN - 1873-295X (E-Journal); 1369-703X (Print) VL - Vol. 73 SP - 29 EP - 37 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Rachinger, Michael A1 - Bauch, Melanie A1 - Strittmatter, Axel A1 - Bongaerts, Johannes A1 - Evers, Stefan A1 - Maurer, Karl-Heinz A1 - Daniel, Rolf A1 - Liebl, Wolfgang A1 - Liesegang, Heiko A1 - Ehrenreich, Armin T1 - Size unlimited markerless deletions by a transconjugative plasmid-system in Bacillus licheniformis JF - Journal of biotechnology Y1 - 2013 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - Vol. 164 IS - Iss. 4 SP - 365 EP - 369 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bandodkar, Amay J. A1 - Molinnus, Denise A1 - Mirza, Omar A1 - Guinovart, Tomas A1 - Windmiller, Joshua R. A1 - Valdes-Ramirez, Gabriela A1 - Andrade, Francisco J. A1 - Schöning, Michael Josef A1 - Wang, Joseph T1 - Epidermal tattoo potentiometric sodium sensors with wireless signal transduction for continuous non-invasive sweat monitoring JF - Biosensors and bioelectronics N2 - This article describes the fabrication, characterization and application of an epidermal temporary-transfer tattoo-based potentiometric sensor, coupled with a miniaturized wearable wireless transceiver, for real-time monitoring of sodium in the human perspiration. Sodium excreted during perspiration is an excellent marker for electrolyte imbalance and provides valuable information regarding an individual's physical and mental wellbeing. The realization of the new skin-worn non-invasive tattoo-like sensing device has been realized by amalgamating several state-of-the-art thick film, laser printing, solid-state potentiometry, fluidics and wireless technologies. The resulting tattoo-based potentiometric sodium sensor displays a rapid near-Nernstian response with negligible carryover effects, and good resiliency against various mechanical deformations experienced by the human epidermis. On-body testing of the tattoo sensor coupled to a wireless transceiver during exercise activity demonstrated its ability to continuously monitor sweat sodium dynamics. The real-time sweat sodium concentration was transmitted wirelessly via a body-worn transceiver from the sodium tattoo sensor to a notebook while the subjects perspired on a stationary cycle. The favorable analytical performance along with the wearable nature of the wireless transceiver makes the new epidermal potentiometric sensing system attractive for continuous monitoring the sodium dynamics in human perspiration during diverse activities relevant to the healthcare, fitness, military, healthcare and skin-care domains. Y1 - 2014 U6 - https://doi.org/10.1016/j.bios.2013.11.039 SN - 1873-4235 (E-Journal); 0956-5663 (Print) VL - 54 SP - 603 EP - 609 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Hirayama, Yuji A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Visualization of enzymatic reaction in a microfluidic channel using chemical imaging sensor JF - Electrochimica acta Y1 - 2013 SN - 1873-3859 (E-Journal); 0013-4686 (Print) SP - Publ. online PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Huck, Christina A1 - Poghossian, Arshak A1 - Bäcker, Matthias A1 - Chaudhuri, S. A1 - Zander, W. A1 - Schubert, J. A1 - Begoyan, V. K. A1 - Buniatyan, V. V. A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Capacitively coupled electrolyte-conductivity sensor based on high-k material of barium strontium titanate JF - Sensors and actuators. B: Chemical Y1 - 2014 U6 - https://doi.org/10.1016/j.snb.2014.02.103 SN - 1873-3077 (E-Journal); 0925-4005 (Print) IS - 198 SP - 102 EP - 109 PB - Elsevier CY - Amsterdam ER - 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 - JOUR A1 - Ribitsch, D. A1 - Karl, W. A1 - Birner-Gruenberger, R. A1 - Gruber, K. A1 - Eiteljoerg, I. A1 - Remler, P. A1 - Wieland, S. A1 - Siegert, Petra A1 - Maurer, Karl-Heinz A1 - Schwab, H. T1 - C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli JF - Journal of biotechnology N2 - Recently, a new alkaline protease named HP70 showing highest homology to extracellular serine proteases of Stenotrophomonas maltophilia and Xanthomonas campestris was found in the course of a metagenome screening for detergent proteases (Niehaus et al., submitted for publication). Attempts to efficiently express the enzyme in common expression hosts had failed. This study reports on the realization of overexpression in Escherichia coli after structural modification of HP70. Modelling of HP70 resulted in a two-domain structure, comprising the catalytic domain and a C-terminal domain which includes about 100 amino acids. On the basis of the modelled structure the enzyme was truncated by deletion of most of the C-terminal domain yielding HP70-C477. This structural modification allowed effective expression of active enzyme using E. coli BL21-Gold as the host. Specific activity of HP70-C477 determined with suc-l-Ala-l-Ala-l-Pro-l-Phe-p-nitroanilide as the substrate was 30 ± 5 U/mg compared to 8 ± 1 U/mg of the native enzyme. HP70-C477 was most active at 40 °C and pH 7–11; these conditions are prerequisite for a potential application as detergent enzyme. Determination of kinetic parameters at 40 °C and pH = 9.5 resulted in KM = 0.23 ± 0.01 mM and kcat = 167.5 ± 3.6 s⁻¹. MS-analysis of peptide fragments obtained from incubation of HP70 and HP70-C477 with insulin B indicated that the C-terminal domain influences the cleavage preferences of the enzyme. Washing experiments confirmed the high potential of HP70-C477 as detergent protease. Y1 - 2010 U6 - https://doi.org/10.1016/j.jbiotec.2010.09.947 SN - 1873-4863 (E-Journal); 0168-1656 (Print) VL - 150 IS - 3 SP - 408 EP - 416 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Dantism, S. A1 - Takenaga, S. A1 - Wagner, P. A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Light-addressable Potentiometric Sensor (LAPS) Combined with Multi-chamber Structures to Investigate the Metabolic Activity of Cells JF - Procedia Engineering N2 - LAPS are field-effect-based potentiometric sensors which are able to monitor analyte concentrations in a spatially resolved manner. Hence, a LAPS sensor system is a powerful device to record chemical imaging of the concentration of chemical species in an aqueous solution, chemical reactions, or the growth of cell colonies on the sensor surface, to record chemical images. In this work, multi-chamber 3D-printed structures made out of polymer (PP-ABS) were combined with LAPS chips to analyse differentially and simultaneously the metabolic activity of Escherichia coli K12 and Chinese hamster ovary (CHO) cells, and the responds of those cells to the addition of glucose solution. Y1 - 2015 U6 - https://doi.org/10.1016/j.proeng.2015.08.647 SN - 1877-7058 N1 - Part of special issue "Eurosensors 2015" VL - 120 SP - 384 EP - 387 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Molinnus, Denise A1 - Bäcker, Matthias A1 - Siegert, Petra A1 - Willenberg, H. A1 - Poghossian, Arshak A1 - Keusgen, M. A1 - Schöning, Michael Josef T1 - Detection of Adrenaline Based on Substrate Recycling Amplification JF - Procedia Engineering N2 - 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. Y1 - 2015 U6 - https://doi.org/10.1016/j.proeng.2015.08.708 SN - 1877-7058 N1 - Eurosensors 2015 VL - 120 SP - 540 EP - 543 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Schusser, Sebastian A1 - Krischer, M. A1 - Molin, D. G. M. A1 - Akker, N. M. S. van den A1 - Bäcker, Matthias A1 - Poghossian, Arshak A1 - Schöning, Michael Josef T1 - Sensor System for in-situ and Real-time Monitoring of Polymer (bio) degradation JF - Procedia Engineering N2 - 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. Y1 - 2015 U6 - https://doi.org/10.1016/j.proeng.2015.08.815 SN - 1877-7058 N1 - Eurosensors 2015 VL - 120 SP - 948 EP - 951 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Pilas, Johanna A1 - Mariano, K. A1 - Keusgen, M. A1 - Selmer, Thorsten A1 - Schöning, Michael Josef T1 - Optimization of an Enzyme-based Multi-parameter Biosensor for Monitoring Biogas Processes JF - Procedia Engineering Y1 - 2015 U6 - https://doi.org/10.1016/j.proeng.2015.08.702 SN - 1877-7058 N1 - Part of special issue "Eurosensors 2015" VL - 120 SP - 532 EP - 535 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Bing, Yu A1 - Wagner, Torsten A1 - Yoshinobu, Tatsuo A1 - Schöning, Michael Josef T1 - Visualization of Defects on a Cultured Cell Layer by Utilizing Chemical Imaging Sensor JF - Procedia Engineering N2 - The chemical imaging sensor is a field-effect sensor which is able to visualize both the distribution of ions (in LAPS mode) and the distribution of impedance (in SPIM mode) inthe sample. In this study, a novel wound-healing assay is proposed, in which the chemical imaging sensor operated in SPIM mode is applied to monitor the defect of a cell layer brought into proximity of the sensing surface.A reduced impedance inside the defect, which was artificially formed ina cell layer, was successfully visualized in a photocurrent image. Y1 - 2015 U6 - https://doi.org/10.1016/j.proeng.2015.08.806 SN - 1877-7058 N1 - Part of special issue "Eurosensors 2015" VL - 120 SP - 936 EP - 939 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bronder, Thomas A1 - Poghossian, Arshak A1 - Scheja, S. A1 - Wu, Chunsheng A1 - Keusgen, M. A1 - Schöning, Michael Josef T1 - Electrostatic Detection of Unlabelled Single- and Double-stranded DNA Using Capacitive Field-effect Devices Functionalized with a Positively Charged Polyelectrolyte Layer JF - Procedia Engineering N2 - Capacitive field-effect electrolyte-insulator-semiconductor sensors consisting of an Al-p-Si-SiO2 structure have been used for the electrical detection of unlabelled single- and double-stranded DNA (dsDNA) molecules by their intrinsic charge. A simple functionalization protocol based on the layer-by-layer (LbL) technique was used to prepare a weak polyelectrolyte/probe-DNA bilayer, followed by the hybridization with complementary target DNA molecules. Due to the flat orientation of the LbL-adsorbed DNA molecules, a high sensor signal has been achieved. In addition, direct label-free detection of in-solution hybridized dsDNA molecules has been studied. Y1 - 2015 U6 - https://doi.org/10.1016/j.proeng.2015.08.710 SN - 1877-7058 N1 - Eurosensors 2015 VL - 120 SP - 544 EP - 547 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Wu, Chunsheng A1 - Poghossian, Arshak A1 - Bronder, Thomas A1 - Schöning, Michael Josef T1 - Sensing of double-stranded DNA molecules by their intrinsic molecular charge using the light-addressable potentiometric sensor JF - Sensors and Actuators B: Chemical N2 - A multi-spot light-addressable potentiometric sensor (LAPS), which belongs to the family of semiconductor field-effect devices, was applied for label-free detection of double-stranded deoxyribonucleic acid (dsDNA) molecules by their intrinsic molecular charge. To reduce the distance between the DNA charge and sensor surface and thus, to enhance the electrostatic coupling between the dsDNA molecules and the LAPS, the negatively charged dsDNA molecules were electrostatically adsorbed onto the gate surface of the LAPS covered with a positively charged weak polyelectrolyte layer of PAH (poly(allylamine hydrochloride)). The surface potential changes in each spot of the LAPS, induced by the layer-by-layer adsorption of a PAH/dsDNA bilayer, were recorded by means of photocurrent-voltage and constant-photocurrent measurements. In addition, the surface morphology of the gate surface before and after consecutive electrostatic adsorption of PAH and dsDNA layers was studied by atomic force microscopy measurements. Moreover, fluorescence microscopy was used to verify the successful adsorption of dsDNA molecules onto the PAH-modified LAPS surface. A high sensor signal of 25 mV was registered after adsorption of 10 nM dsDNA molecules. The lower detection limit is down to 0.1 nM dsDNA. The obtained results demonstrate that the PAH-modified LAPS device provides a convenient and rapid platform for the direct label-free electrical detection of in-solution hybridized dsDNA molecules. KW - Layer-by-layer adsorption KW - Poly(allylamine hydrochloride) KW - Label-free detection KW - DNA biosensor KW - LAPS KW - Field effect Y1 - 2016 U6 - https://doi.org/10.1016/j.snb.2016.02.004 SN - 0925-4005 IS - 229 SP - 506 EP - 512 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Handtke, Stefan A1 - Volland, Sonja A1 - Methling, Karen A1 - Albrecht, Dirk A1 - Becher, Dörte A1 - Nehls, Jenny A1 - Bongaerts, Johannes A1 - Maurer, Karl-Heinz A1 - Lalk, Michael A1 - Liesegang, Heiko A1 - Voigt, Birgit A1 - Daniel, Rolf A1 - Hecker, Michael T1 - Cell physiology of the biotechnological relevant bacterium Bacillus pumilus - An omics-based approach JF - Journal of Biotechnology N2 - Members of the species Bacillus pumilus get more and more in focus of the biotechnological industry as potential new production strains. Based on exoproteome analysis, B. pumilus strain Jo2, possessing a high secretion capability, was chosen for an omics-based investigation. The proteome and metabolome of B. pumilus cells growing either in minimal or complex medium was analyzed. In total, 1542 proteins were identified in growing B. pumilus cells, among them 1182 cytosolic proteins, 297 membrane and lipoproteins and 63 secreted proteins. This accounts for about 43% of the 3616 proteins encoded in the B. pumilus Jo2 genome sequence. By using GC–MS, IP-LC/MS and H NMR methods numerous metabolites were analyzed and assigned to reconstructed metabolic pathways. In the genome sequence a functional secretion system including the components of the Sec- and Tat-secretion machinery was found. Analysis of the exoproteome revealed secretion of about 70 proteins with predicted secretion signals. In addition, selected production-relevant genome features such as restriction modification systems and NRPS clusters of B. pumilus Jo2 are discussed. Y1 - 2014 U6 - https://doi.org/10.1016/j.jbiotec.2014.08.028 SN - 1873-4863 (E-Journal); 0168-1656 (Print) IS - 192(A) SP - 204 EP - 214 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Guo, Yuanyuan A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Device simulation of the light-addressable potentiometric sensor for the investigation of the spatial resolution JF - Sensors and actuators B: Chemical N2 - As a semiconductor-based electrochemical sensor, the light-addressable potentiometric sensor (LAPS) can realize two dimensional visualization of (bio-)chemical reactions at the sensor surface addressed by localized illumination. Thanks to this imaging capability, various applications in biochemical and biomedical fields are expected, for which the spatial resolution is critically significant. In this study, therefore, the spatial resolution of the LAPS was investigated in detail based on the device simulation. By calculating the spatiotemporal change of the distributions of electrons and holes inside the semiconductor layer in response to a modulated illumination, the photocurrent response as well as the spatial resolution was obtained as a function of various parameters such as the thickness of the Si substrate, the doping concentration, the wavelength and the intensity of illumination. The simulation results verified that both thinning the semiconductor substrate and increasing the doping concentration could improve the spatial resolution, which were in good agreement with known experimental results and theoretical analysis. More importantly, new findings of interests were also obtained. As for the dependence on the wavelength of illumination, it was found that the known dependence was not always the case. When the Si substrate was thick, a longer wavelength resulted in a higher spatial resolution which was known by experiments. When the Si substrate was thin, however, a longer wavelength of light resulted in a lower spatial resolution. This finding was explained as an effect of raised concentration of carriers, which reduced the thickness of the space charge region. The device simulation was found to be helpful to understand the relationship between the spatial resolution and device parameters, to understand the physics behind it, and to optimize the device structure and measurement conditions for realizing higher performance of chemical imaging systems. Y1 - 2014 U6 - https://doi.org/10.1016/j.snb.2014.08.016 SN - 1873-3077 (E-Journal); 0925-4005 (Print) VL - 204 SP - 659 EP - 665 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Reisert, Steffen A1 - Geissler, H. A1 - Weiler, C. A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - Multiple sensor-type system for monitoring the microbicidal effectiveness of aseptic sterilisation processes JF - Food control N2 - The present work describes a novel multiple sensor-type system for the real-time analysis of aseptic sterilisation processes employing gaseous hydrogen peroxide (H2O2) as a sterilant. The inactivation kinetics of Bacillus atrophaeus by gaseous H2O2 have been investigated by means of a methodical calibration experiment, taking into account the process variables H2O2 concentration, humidity and gas temperature. It has been found that the microbicidal effectiveness at H2O2 concentrations above 2% v/v is largely determined by the concentration itself, while at lower H2O2 concentrations, the gas temperature and humidity play a leading role. Furthermore, the responses of different types of gas sensors towards the influencing factors of the sterilisation process have been analysed within the same experiment. Based on a correlation established between the inactivation kinetics and the sensor responses, a calorimetric H2O2 sensor and a metal-oxide semiconductor (MOX) sensor have been identified as possible candidates for monitoring the microbicidal effectiveness of aseptic sterilisation processes employing gaseous H2O2. Therefore, two linear models that describe the relationship between sensor response and microbicidal effectiveness have been proposed. Y1 - 2015 U6 - https://doi.org/10.1016/j.foodcont.2014.07.063 SN - 1873-7129 (E-Journal); 0956-7135 (Print) VL - 47 SP - 615 EP - 622 ER - TY - JOUR A1 - Schusser, Sebastian A1 - Poghossian, Arshak A1 - Bäcker, Matthias A1 - Krischer, M. A1 - Leinhos, Marcel A1 - Wagner, P. A1 - Schöning, Michael Josef T1 - An application of field-effect sensors for in-situ monitoring of degradation of biopolymers JF - Sensors and actuators B: Chemical N2 - 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 500 nm 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). Y1 - 2015 U6 - https://doi.org/10.1016/j.snb.2014.10.058 SN - 1873-3077 (E-Journal); 0925-4005 (Print) VL - 207, Part B SP - 954 EP - 959 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Yoshinobu, Tatsuo A1 - Miyamoto, Ko-ichiro A1 - Wagner, Torsten A1 - Schöning, Michael Josef T1 - Recent developments of chemical imaging sensor systems based on the principle of the light-addressable potentiometric sensor JF - Sensors and actuators B: Chemical N2 - The light-addressable potentiometric sensor (LAPS) is an electrochemical sensor with a field-effect structure to detect the variation of the Nernst potential at its sensor surface, the measured area on which is defined by illumination. Thanks to this light-addressability, the LAPS can be applied to chemical imaging sensor systems, which can visualize the two-dimensional distribution of a particular target ion on the sensor surface. Chemical imaging sensor systems are expected to be useful for analysis of reaction and diffusion in various electrochemical and biological samples. Recent developments of LAPS-based chemical imaging sensor systems, in terms of the spatial resolution, measurement speed, image quality, miniaturization and integration with microfluidic devices, are summarized and discussed. Y1 - 2015 U6 - https://doi.org/10.1016/j.snb.2014.09.002 SN - 1873-3077 (E-Journal); 0925-4005 (Print) VL - 207, Part B SP - 926 EP - 932 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Murib, M. S. A1 - Yeap, W. S. A1 - Martens, D. A1 - Liu, X. A1 - Bienstman, P. A1 - Fahlman, M. A1 - Schöning, Michael Josef A1 - Michiels, L. A1 - Haenen, K. A1 - Serpengüzel, A. A1 - Wagner, Patrick T1 - Photonic studies on polymer-coated sapphire-spheres : a model system for biological ligands JF - Sensors and actuators A: Physical N2 - In this study we show an optical biosensor concept, based on elastic light scattering from sapphire microspheres. Transmitted and elastic scattering intensity of the microspheres (radius 500 μm, refractive index 1.77) on an optical fiber half coupler is analyzed at 1510 nm. The 0.43 nm angular mode spacing of the resonances is comparable to the angular mode spacing value estimated using the optical size of the microsphere. The spectral linewidths of the resonances are in the order of 0.01 nm, which corresponds to quality factors of approximately 105. A polydopamine layer is used as a functionalizing agent on sapphire microspherical resonators in view of biosensor implementation. The varying layer thickness on the microsphere is determined as a function of the resonance wavelength shift. It is shown that polymer functionalization has a minor effect on the quality factor. This is a promising step toward the development of an optical biosensor. Y1 - 2015 U6 - https://doi.org/10.1016/j.sna.2014.11.024 SN - 1873-3069 (E-Journal); 0924-4247 (Print) VL - 222 SP - 212 EP - 219 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, K. A1 - Seki, K. A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, T. T1 - Enhancement of the spatial resolution of the chemical imaging sensor by a hybrid fiber-optic illumination JF - Procedia Engineering N2 - The chemical imaging sensor, which is based on the principle of the light-addressable potentiometric sensor (LAPS), is a powerful tool to visualize the spatial distribution of chemical species on the sensor surface. The spatial resolution of this sensor depends on the diffusion of photocarriers excited by a modulated light. In this study, a novel hybrid fiber-optic illumination was developed to enhance the spatial resolution. It consists of a modulated light probe to generate a photocurrent signal and a ring of constant light, which suppresses the lateral diffusion of minority carriers excited by the modulated light. It is demonstrated that the spatial resolution was improved from 92 μm to 68 μm. Y1 - 2014 U6 - https://doi.org/10.1016/j.proeng.2014.11.563 SN - 1877-7058 N1 - EUROSENSORS 2014 ; European Conference on Solid-State Transducers <28, 2014> VL - 87 SP - 612 EP - 615 PB - Elsevier CY - Amsterdam 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 - Oberländer, Jan A1 - Kirchner, Patrick A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Strategies in developing thin-film sensors for monitoring aseptic food processes : Theoretical considerations and investigations of passivation materials JF - Electrochimica Acta N2 - The sterilization of packages in aseptic food processes is highly significant to maintain a consumer-safe product with extended shelf-life. Today, the sterilization of food packages is predominantly accomplished by gaseous hydrogen peroxide (H2O2) in combination with heat. In order to monitor this sterilization process, calorimetric gas sensors as differential set-up of two platinum temperature sensors representing a catalytically active (additionally deposition of MnO2) and a passive segment have been recently developed. The temperature rise of the exothermic decomposition serves as an indicator of the present H2O2 concentration. In the present work, a theoretical approach considering the sensor’s thermochemistry and physical transport phenomena was formulated to evaluate the temperature rise based on the energy content of gaseous H2O2. In a further part of this work, three polymers have been analyzed with respect to their application as passivation materials. The examined polymers are photoresist SU-8, perfluoroalkoxy (PFA) and fluorinated ethylene propylene (FEP). Thermal analyses by means of differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) have been conducted to determine the operation limits of the polymers. The overall chemical resistance and stability of the polymers against the harsh environmental conditions during the sterilization process have been examined by attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR). Y1 - 2015 U6 - https://doi.org/10.1016/j.electacta.2015.06.126 SN - 0013-4686 VL - 183 SP - 130 EP - 136 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Murib, M. S. A1 - Yeap, W. S. A1 - Eurlings, Y. A1 - Grinsven, B. van A1 - Boyen, H.-G. A1 - Conings, B. A1 - Michiels, L. A1 - Ameloot, M. A1 - Carleer, R. A1 - Warmer, J. A1 - Kaul, P. A1 - Haenen, K. A1 - Schöning, Michael Josef A1 - Ceuninck, W. de A1 - Wagner, P. T1 - Heat-transfer based characterization of DNA on synthetic sapphire chips JF - Sensors and Actuators B: Chemical N2 - In this study, we show that synthetic sapphire (Al₂O₃), an established implant material, can also serve as a platform material for biosensors comparable to nanocrystalline diamond. Sapphire chips, beads, and powder were first modified with (3-aminopropyl) triethoxysilane (APTES), followed by succinic anhydride (SA), and finally single-stranded probe DNA was EDC coupled to the functionalized layer. The presence of the APTES-SA layer on sapphire powders was confirmed by thermogravimetric analyis and Fourier-transform infrared spectroscopy. Using planar sapphire chips as substrates and X-ray photoelectron spectroscopy (XPS) as surface-sensitive tool, the sequence of individual layers was analyzed with respect to their chemical state, enabling the quantification of areal densities of the involved molecular units. Fluorescence microscopy was used to demonstrate the hybridization of fluorescently tagged target DNA to the probe DNA, including denaturation- and re-hybridization experiments. Due to its high thermal conductivity, synthetic sapphire is especially suitable as a chip material for the heat-transfer method, which was employed to distinguish complementary- and non-complementary DNA duplexes containing single-nucleotide polymorphisms. These results indicate that it is possible to detect mutations electronically with a chemically resilient and electrically insulating chip material. Y1 - 2016 U6 - https://doi.org/10.1016/j.snb.2016.02.027 SN - 0925-4005 VL - 230 IS - 230 SP - 260 EP - 271 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamato, Ko-ichiro A1 - Sakakita, Sakura A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Application of chemical imaging sensor to in-situ pH imaging in the vicinity of a corroding metal surface JF - Electrochimica Acta N2 - The chemical imaging sensor was applied to in-situ pH imaging of the solution in the vicinity of a corroding surface of stainless steel under potentiostatic polarization. A test piece of polished stainless steel was placed on the sensing surface leaving a narrow gap filled with artificial seawater and the stainless steel was corroded under polarization. The pH images obtained during polarization showed correspondence between the region of lower pH and the site of corrosion. It was also found that the pH value in the gap became as low as 2 by polarization, which triggered corrosion. Y1 - 2015 U6 - https://doi.org/10.1016/j.electacta.2015.07.184 SN - 0013-4686 VL - 183 SP - 137 EP - 142 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Miyamoto, Ko-ichiro A1 - Yu, Bing A1 - Isoda, Hiroko A1 - Wagner, Torsten A1 - Schöning, Michael Josef A1 - Yoshinobu, Tatsuo T1 - Visualization of the recovery process of defects in a cultured cell layer by chemical imaging sensor JF - Sensors and Actuators B: Chemical N2 - The chemical imaging sensor is a field-effect sensor which is able to visualize both the distribution of ions (in LAPS mode) and the distribution of impedance (in SPIM mode) in the sample. In this study, a novel cell assay is proposed, in which the chemical imaging sensor operated in SPIM mode is applied to monitor the recovery of defects in a cell layer brought into proximity of the sensing surface. A reduced impedance at a defect formed artificially in a cell layer was successfully visualized in a photocurrent image. The cell layer was cultured over two weeks, during which the temporal change of the photocurrent distribution corresponding to the recovery of the defect was observed. Y1 - 2016 U6 - https://doi.org/10.1016/j.snb.2016.04.018 SN - 0925-4005 VL - 236 SP - 965 EP - 969 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Bäcker, Matthias A1 - Koch, Claudia A1 - Eiben, Sabine A1 - Geiger, Fania A1 - Eber, Fabian A1 - Gliemann, Hartmut A1 - Poghossian, Arshak A1 - Wege, Christina A1 - Schöning, Michael Josef T1 - Tobacco mosaic virus as enzyme nanocarrier for electrochemical biosensors JF - Sensors and Actuators B: Chemical N2 - 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. Y1 - 2017 U6 - https://doi.org/10.1016/j.snb.2016.07.096 SN - 0925-4005 VL - 238 SP - 716 EP - 722 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Wagner, Torsten A1 - Vornholt, Wolfgang A1 - Werner, Frederik A1 - Yoshinobu, Tatsuo A1 - Miyamoto, Ko-Ichiro A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Light-addressable potentiometric sensor (LAPS) combined with magnetic beads for pharmaceutical screening JF - Physics in medicine N2 - The light-addressable potentiometric sensor (LAPS) has the unique feature to address different regions of a sensor surface without the need of complex structures. Measurements at different locations on the sensor surface can be performed in a common analyte solution, which distinctly simplifies the fluidic set-up. However, the measurement in a single analyte chamber prevents the application of different drugs or different concentrations of a drug to each measurement spot at the same time as in the case of multi-reservoir-based set-ups. In this work, the authors designed a LAPS-based set-up for cell culture screening that utilises magnetic beads loaded with the endotoxin (lipopolysaccharides, LPS), to generate a spatially distributed gradient of analyte concentration. Different external magnetic fields can be adjusted to move the magnetic beads loaded with a specific drug within the measurement cell. By recording the metabolic activities of a cell layer cultured on top of the LAPS surface, this work shows the possibility to apply different concentrations of a sample along the LAPS measurement spots within a common analyte solution. Y1 - 2016 U6 - https://doi.org/10.1016/j.phmed.2016.03.001 SN - 2352-4510 VL - 2016 IS - 1 SP - 2 EP - 7 ER - TY - JOUR A1 - Molinnus, Denise A1 - Sorich, Maren A1 - Bartz, Alexander A1 - Siegert, Petra A1 - Willenberg, Holger S. A1 - Lisdat, Fred A1 - Poghossian, Arshak A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Towards an adrenaline biosensor based on substrate recycling amplification in combination with an enzyme logic gate JF - Sensors and Actuators B: Chemical N2 - An amperometric biosensor using a substrate recycling principle was realized for the detection of low adrenaline concentrations (1 nM) by measurements in phosphate buffer and Ringer’s solution at pH 6.5 and pH 7.4, respectively. In proof-of-concept experiments, a Boolean logic-gate principle has been applied to develop a digital adrenaline biosensor based on an enzyme AND logic gate. The obtained results demonstrate that the developed digital biosensor is capable for a rapid qualitative determination of the presence/absence of adrenaline in a YES/NO statement. Such digital biosensor could be used in clinical diagnostics for the control of a correct insertion of a catheter in the adrenal veins during adrenal venous-sampling procedure. Y1 - 2016 U6 - https://doi.org/10.1016/j.snb.2016.06.064 SN - 0925-4005 VL - 237 SP - 190 EP - 195 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Molinnus, Denise A1 - Hardt, G. A1 - Käver, L. A1 - Willenberg, H.S. A1 - Kröger, J.-C. A1 - Poghossian, Arshak A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Chip-based biosensor for the detection of low adrenaline concentrations to support adrenal venous sampling JF - Sensor and Actuators B: Chemical N2 - A chip-based amperometric biosensor referring on using the bioelectrocatalytical amplification principle for the detection of low adrenaline concentrations is presented. The adrenaline biosensor has been prepared by modification of a platinum thin-film electrode with an enzyme membrane containing the pyrroloquinoline quinone-dependent glucose dehydrogenase and glutaraldehyde. Measuring conditions such as temperature, pH value, and glucose concentration have been optimized to achieve a high sensitivity and a low detection limit of about 1 nM adrenaline measured in phosphate buffer at neutral pH value. The response of the biosensor to different catecholamines has also been proven. Long-term stability of the adrenaline biosensor has been studied over 10 days. In addition, the biosensor has been successfully applied for adrenaline detection in human blood plasma for future biomedical applications. Furthermore, preliminary experiments have been carried to detect the adrenaline-concentration difference measured in peripheral blood and adrenal venous blood, representing the adrenal vein sampling procedure of a physician. Y1 - 2018 U6 - https://doi.org/10.1016/j.snb.2018.05.136 SN - 0925-4005 VL - 272 SP - 21 EP - 27 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Aboulnaga, E. A. A1 - Zou, H. A1 - Selmer, Thorsten A1 - Xian, M. T1 - Development of a plasmid-based, tunable, tolC-derived expression system for application in Cupriavidus necator H16 JF - Journal of Biotechnology N2 - Cupriavidus necator H16 gains increasing attention in microbial research and biotechnological application due to its diverse metabolic features. Here we present a tightly controlled gene expression system for C. necator including the pBBR1-vector that contains hybrid promoters originating from C. necator native tolC-promoter in combination with a synthetic tetO-operator. The expression of the reporter gene from these plasmids relies on the addition of the exogenous inducer doxycycline (dc). The novel expression system offers a combination of advantageous features as; (i) high and dose-dependent recombinant protein production, (ii) tight control with a high dynamic range (On/Off ratio), which makes it applicable for harmful pathways or for toxic protein production, (iii) comparable cheap inducer (doxycycline, dc), (iv) effective at low inducer concentration, that makes it useful for large scale application, (v) rapid, diffusion controlled induction, and (vi) the inducer does not interfere within the cell metabolism. As applications of the expression system in C. necator H16, the growth ability on glycerol was enhanced by constitutively expressing the E. coli glpk gene-encoding for glycerol kinase. Likewise, we used the system to overcome the expression toxicity of mevalonate pathway in C. necator H16. With this system, the mevalonate-genes were successfully introduced in the host and the recombinant strains could produce about 200 mg/l mevalonate. Y1 - 2018 U6 - https://doi.org/10.1016/j.jbiotec.2018.03.007 SN - 0168-1656 VL - 274 SP - 15 EP - 27 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Molinnus, Denise A1 - Muschallik, Lukas A1 - Gonzalez, Laura Osorio A1 - Bongaerts, Johannes A1 - Wagner, Torsten A1 - Selmer, Thorsten A1 - Siegert, Petra A1 - Keusgen, Michael A1 - Schöning, Michael Josef T1 - Development and characterization of a field-effect biosensor for the detection of acetoin JF - Biosensors and Bioelectronics N2 - A capacitive electrolyte-insulator-semiconductor (EIS) field-effect biosensor for acetoin detection has been presented for the first time. The EIS sensor consists of a layer structure of Al/p-Si/SiO₂/Ta₂O₅/enzyme acetoin reductase. The enzyme, also referred to as butane-2,3-diol dehydrogenase from B. clausii DSM 8716T, has been recently characterized. The enzyme catalyzes the (R)-specific reduction of racemic acetoin to (R,R)- and meso-butane-2,3-diol, respectively. Two different enzyme immobilization strategies (cross-linking by using glutaraldehyde and adsorption) have been studied. Typical biosensor parameters such as optimal pH working range, sensitivity, hysteresis, linear concentration range and long-term stability have been examined by means of constant-capacitance (ConCap) mode measurements. Furthermore, preliminary experiments have been successfully carried out for the detection of acetoin in diluted white wine samples. Y1 - 2018 U6 - https://doi.org/10.1016/j.bios.2018.05.023 VL - 115 SP - 1 EP - 6 PB - Elsevier CY - Amsterdam ER -