@article{KochPoghossianSchoeningetal.2018, author = {Koch, Claudia and Poghossian, Arshak and Sch{\"o}ning, Michael Josef and Wege, Christian}, title = {Penicillin Detection by Tobacco Mosaic Virus-Assisted Colorimetric Biosensors}, series = {Nanotheranostics}, volume = {2}, journal = {Nanotheranostics}, number = {2}, publisher = {Ivyspring}, address = {Sydney}, issn = {2206-7418}, doi = {10.7150/ntno.22114}, pages = {184 -- 196}, year = {2018}, abstract = {The presentation of enzymes on viral scaffolds has beneficial effects such as an increased enzyme loading and a prolonged reusability in comparison to conventional immobilization platforms. Here, we used modified tobacco mosaic virus (TMV) nanorods as enzyme carriers in penicillin G detection for the first time. Penicillinase enzymes were conjugated with streptavidin and coupled to TMV rods by use of a bifunctional biotin-linker. Penicillinase-decorated TMV particles were characterized extensively in halochromic dye-based biosensing. Acidometric analyte detection was performed with bromcresol purple as pH indicator and spectrophotometry. The TMV-assisted sensors exhibited increased enzyme loading and strongly improved reusability, and higher analysis rates compared to layouts without viral adapters. They extended the half-life of the sensors from 4 - 6 days to 5 weeks and thus allowed an at least 8-fold longer use of the sensors. Using a commercial budget-priced penicillinase preparation, a detection limit of 100 µM penicillin was obtained. Initial experiments also indicate that the system may be transferred to label-free detection layouts.}, language = {en} } @article{BegingMlynekHataihimakuletal.2010, author = {Beging, Stefan and Mlynek, Daniela and Hataihimakul, Sudkanung and Poghossian, Arshak and Baldsiefen, Gerhard and Busch, Heinz and Laube, Norbert and Kleinen, Lisa and Sch{\"o}ning, Michael Josef}, title = {Field-effect calcium sensor for the determination of the risk of urinary stone formation}, series = {Sensors and Actuators B: Chemical}, volume = {144}, journal = {Sensors and Actuators B: Chemical}, number = {2}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2008.12.012}, pages = {374 -- 379}, year = {2010}, abstract = {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.}, language = {en} } @article{ReisertHenkelSchneideretal.2010, author = {Reisert, Steffen and Henkel, Hartmut and Schneider, Andreas and Sch{\"a}fer, Daniel and Friedrich, Peter and Berger, J{\"o}rg and Sch{\"o}ning, Michael Josef}, title = {Development of a handheld sensor system for the online measurement of hydrogen peroxide in aseptic filling systems}, series = {Physica Status Solidi (A)}, volume = {207}, journal = {Physica Status Solidi (A)}, number = {4}, publisher = {Wiley-VCH}, address = {Berlin}, issn = {1862-6300}, doi = {10.1002/pssa.200983304}, pages = {913 -- 918}, year = {2010}, abstract = {A handheld sensor system for the online measurement of hydrogen peroxide (H2O2) in aseptic sterilisation processes has been developed. It is based on a calorimetric-type gas sensor that consists of a differential set-up of two temperature sensors, of which one is catalytically activated and the second one is passivated and used as reference. The sensor principle relies in detecting a rise in temperature on the active sensor due to the exothermic reaction of H2O2 on the catalytic surface. To characterise the sensor system towards H2O2 sensitivity and other influencing factors, measurements have been carried out both at an experimental set-up and a manufacturer's sterilisation machine. Physical sensor characterisation was done by means of the optical microscopy.}, language = {en} } @article{MiyamotoSugawaraKanohetal.2010, author = {Miyamoto, Ko-ichiro and Sugawara, Yuri and Kanoh, Shin´ichiro and Yoshinobu, Tatsuo and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Image correction method for the chemical imaging sensor}, series = {Sensors and Actuators B: Chemical}, volume = {144}, journal = {Sensors and Actuators B: Chemical}, number = {2}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2008.10.069}, pages = {344 -- 348}, year = {2010}, abstract = {The chemical imaging sensor is a semiconductor-based chemical sensor that can visualize the spatial distribution of chemical species. For the practical application of this sensor, artifacts in the chemical images due to defects of the semiconductor substrate and contamination of the sensing surface etc. have been a major problem. An image correction method was developed to eliminate the influence of nonuniformity of individual sensor plate.}, language = {en} } @article{AbouzarPoghossianSiqueiraetal.2010, author = {Abouzar, Maryam H. and Poghossian, Arshak and Siqueira, Jos{\´e} R. Jr. and Oliveira, Osvaldo N. Jr. and Moritz, Werner and Sch{\"o}ning, Michael Josef}, title = {Capacitive electrolyte-insulator-semiconductor structures functionalised with a polyelectrolyte/enzyme multilayer: New strategy for enhanced field-effect biosensing}, series = {Physica Status Solidi (A)}, volume = {207}, journal = {Physica Status Solidi (A)}, number = {4}, publisher = {Wiley-VCH}, address = {Berlin}, issn = {1862-6300}, doi = {10.1002/pssa.200983317}, pages = {884 -- 890}, year = {2010}, abstract = {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\%).}, language = {en} } @article{PoghossianJablonskiKochetal.2018, author = {Poghossian, Arshak and Jablonski, Melanie and Koch, Claudia and Bronder, Thomas and Rolka, David and Wege, Christina and Sch{\"o}ning, Michael Josef}, title = {Field-effect biosensor using virus particles as scaffolds for enzyme immobilization}, series = {Biosensors and Bioelectronics}, volume = {110}, journal = {Biosensors and Bioelectronics}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0956-5663}, doi = {10.1016/j.bios.2018.03.036}, pages = {168 -- 174}, year = {2018}, abstract = {A field-effect biosensor employing tobacco mosaic virus (TMV) particles as scaffolds for enzyme immobilization is presented. Nanotubular TMV scaffolds allow a dense immobilization of precisely positioned enzymes with retained activity. To demonstrate feasibility of this new strategy, a penicillin sensor has been developed by coupling a penicillinase with virus particles as a model system. The developed field-effect penicillin biosensor consists of an Al-p-Si-SiO₂-Ta₂O₅-TMV structure and has been electrochemically characterized in buffer solutions containing different concentrations of penicillin G. In addition, the morphology of the biosensor surface with virus particles was characterized by scanning electron microscopy and atomic force microscopy methods. The sensors possessed a high penicillin sensitivity of ~ 92 mV/dec in a nearly-linear range from 0.1 mM to 10 mM, and a low detection limit of about 50 µM. The long-term stability of the penicillin biosensor was periodically tested over a time period of about one year without any significant loss of sensitivity. The biosensor has also been successfully applied for penicillin detection in bovine milk samples.}, language = {en} } @article{SchoeningKirchnerNgetal.2010, author = {Sch{\"o}ning, Michael Josef and Kirchner, Patrick and Ng, Yue Ann and Spelthahn, Heiko and Schneider, Andreas and Henkel, Hartmut and Friedrich, Peter and Kolstad, Jens and Berger, J{\"o}rg and Keusgen, Michael}, title = {Gas sensor investigation based on a catalytically activated thin-film thermopile for H2O2 detection}, series = {Physica Status Solidi (A)}, volume = {207}, journal = {Physica Status Solidi (A)}, number = {4}, publisher = {Wiley-VCH}, address = {Berlin}, issn = {1862-6300}, doi = {10.1002/pssa.200983309}, pages = {787 -- 792}, year = {2010}, abstract = {In aseptic filling systems, hydrogen peroxide vapour is commonly used for the reduction of microbial contaminations in carton packages. In this process, the germicidal efficiency of the vapour depends especially on the H₂O₂ concentration. To monitor the H₂O₂ concentration, a calorimetric H₂O₂ gas sensor based on a catalytically activated thin-film thermopile is investigated. Two different sensor layouts, namely a circular and a linear form, as well as two various material pairs such as tungsten/nickel and gold/nickel, have been examined for the realization of a thin-film thermopile. Additionally, manganese oxide and palladium particles have been compared as responsive catalysts towards H₂O₂. The thin-film sensors have been investigated at various H₂O₂ concentrations, gas temperatures and flow rates.}, language = {en} } @article{HuckPoghossianBaeckeretal.2014, author = {Huck, Christina and Poghossian, Arshak and B{\"a}cker, Matthias and Chaudhuri, S. and Zander, W. and Schubert, J. and Begoyan, Vardges K. and Buniatyan, V. V. and Wagner, P. and Sch{\"o}ning, Michael Josef}, title = {Capacitively coupled electrolyte-conductivity sensor based on high-k material of barium strontium titanate}, series = {Sensors and actuators. B: Chemical}, journal = {Sensors and actuators. B: Chemical}, number = {198}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-3077 (E-Journal); 0925-4005 (Print)}, doi = {10.1016/j.snb.2014.02.103}, pages = {102 -- 109}, year = {2014}, language = {en} } @article{BuniatyanAbouzarMartirosyanetal.2010, author = {Buniatyan, Vahe V. and Abouzar, Maryam H. and Martirosyan, Norayr W. and Schubert, J{\"u}rgen and Gevorgian, Spartak and Sch{\"o}ning, Michael Josef and Poghossian, Arshak}, title = {pH-sensitive properties of barium strontium titanate (BST) thin films prepared by pulsed laser deposition technique}, series = {Physica Status Solidi (A)}, volume = {207}, journal = {Physica Status Solidi (A)}, number = {4}, publisher = {Wiley-VCH}, address = {Berlin}, issn = {1862-6300}, doi = {10.1002/pssa.200983310}, pages = {824 -- 830}, year = {2010}, abstract = {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.}, language = {en} } @article{TurekHeidenGuoetal.2010, author = {Turek, Monik and Heiden, Wolfgang and Guo, Sharon and Riesen, Alfred and Schubert, J{\"u}rgen and Zander, Willi and Kr{\"u}ger, Peter and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Simultaneous detection of cyanide and heavy metals for environmental analysis by means of µISEs}, series = {Physica Status Solidi (A)}, volume = {207}, journal = {Physica Status Solidi (A)}, number = {4}, publisher = {Wiley-VCH}, address = {Berlin}, issn = {1862-6300}, doi = {10.1002/pssa.200983303}, pages = {817 -- 823}, year = {2010}, abstract = {In environmental analysis, cyanide and heavy metals play an important role, because these substances are highly toxic for biological systems. They can lead to chronic and acute diseases. Due to the chemical properties of cyanide it is frequently used for industrial processes such as extraction of silver and gold. Heavy metals can be found as trace elements in nature and are often applied in industries e.g., galvanization processes. Up to now, cyanide and heavy metals can be detected by several sensors separately and their detection is often limited to laboratory investigations. In this publication, with regard to an in situ analysis, a new miniaturized silicon-based sensor system for the simultaneous detection of cyanide and heavy metals in aqueous solutions is presented that is based on chalcogenide glass-based micro ion-selective electrodes (µISEs). The µISEs are incorporated into a specially designed measuring system for the simultaneous detection of heavy metals and cyanide in solutions and validated by simultaneous measurements of Cu2+- and CN--ions, Cd2+- and CN-- ions and Pb2+- and CN--ions. The particular sensor system has shown good sensor properties in the µ-molar ion-concentration range. For simultaneous measurements in complex heavy metal and cyanide solutions an intelligent software using fuzzy logic is discussed.}, language = {en} } @article{SiqueiraBaeckerPoghossianetal.2010, author = {Siqueira, Jos{\´e} R. Jr. and B{\"a}cker, Matthias and Poghossian, Arshak and Zucolotto, Valtencir and Oliveira, Osvaldo N. Jr. and Sch{\"o}ning, Michael Josef}, title = {Associating biosensing properties with the morphological structure of multilayers containing carbon nanotubes on field-effect devices}, series = {Physica Status Solidi (A)}, volume = {207}, journal = {Physica Status Solidi (A)}, number = {4}, publisher = {Wiley-VCH}, address = {Berlin}, issn = {1862-6300}, doi = {10.1002/pssa.200983301}, pages = {781 -- 786}, year = {2010}, abstract = {The control of molecular architecture provided by the layer-by-layer (LbL) technique has led to enhanced biosensors, in which advantageous features of distinct materials can be combined. Full optimization of biosensing performance, however, is only reached if the film morphology is suitable for the principle of detection of a specific biosensor. In this paper, we report a detailed morphology analysis of LbL films made with alternating layers of single-walled carbon nanotubes (SWNTs) and polyamidoamine (PAMAM) dendrimers, which were then covered with a layer of penicillinase (PEN). An optimized performance to detect penicillin G was obtained with 6-bilayer SWNT/PAMAM LbL films deposited on p-Si-SiO2-Ta2O5 chips, used in biosensors based on a capacitive electrolyte-insulator-semiconductor (EIS) and a light-addressable potentiometric sensor (LAPS) structure, respectively. Field-emission scanning electron microscopy (FESEM) and atomic force microscopy (AFM) images indicated that the LbL films were porous, with a large surface area due to interconnection of SWNT into PAMAM layers. This morphology was instrumental for the adsorption of a larger quantity of PEN, with the resulting LbL film being highly stable. The experiments to detect penicillin were performed with constant-capacitance (ConCap) and constant-current (CC) measurements for EIS and LAPS sensors, respectively, which revealed an enhanced detection signal and sensitivity of ca. 100 mV/decade for the field-effect sensors modified with the PAMAM/SWNT LbL film. It is concluded that controlling film morphology is essential for an enhanced performance of biosensors, not only in terms of sensitivity but also stability and response time.}, language = {en} } @article{RibitschHeumannKarletal.2012, author = {Ribitsch, Doris and Heumann, Sonja and Karl, Wolfgang and Gerlach, Jochen and Leber, R. and Birner-Gruenberger, R. and Gruber, K. and Eiteljoerg, I. and Remler, Peter and Siegert, Petra and Lange, J. and Maurer, Karl-Heinz and Berg, G. and Guebitz, G. M. and Schwab, H.}, title = {Extracellular serine proteases from Stenotrophomonas maltophilia: Screening, isolation and heterologous expression in E. coli}, series = {Journal of biotechnology}, volume = {157}, journal = {Journal of biotechnology}, number = {1}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2011.09.025}, pages = {140 -- 147}, year = {2012}, abstract = {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.}, language = {en} } @article{RibitschKarlBirnerGruenbergeretal.2010, author = {Ribitsch, Doris and Karl, Wolfgang and Birner-Gruenberger, R. and Gruber, K. and Eiteljoerg, I. and Remler, Peter and Wieland, S. and Siegert, Petra and Maurer, Karl-Heinz and Schwab, H.}, title = {C-terminal truncation of a metagenome-derived detergent protease for effective expression in E. coli}, series = {Journal of biotechnology}, volume = {150}, journal = {Journal of biotechnology}, number = {3}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1873-4863 (E-Journal); 0168-1656 (Print)}, doi = {10.1016/j.jbiotec.2010.09.947}, pages = {408 -- 416}, year = {2010}, abstract = {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.}, language = {en} }