@inproceedings{JablonskiKochBronderetal.2017, author = {Jablonski, Melanie and Koch, Claudia and Bronder, Thomas and Poghossian, Arshak and Wege, Christina and Sch{\"o}ning, Michael Josef}, title = {Field-Effect Biosensors Modified with Tobacco Mosaic Virus Nanotubes as Enzyme Nanocarrier}, series = {MDPI Proceeding}, volume = {1}, booktitle = {MDPI Proceeding}, number = {4}, doi = {10.3390/proceedings1040505}, pages = {4}, year = {2017}, language = {en} } @inproceedings{OberlaenderArreolaHansenetal.2017, author = {Oberl{\"a}nder, Jan and Arreola, Julio and Hansen, Christina and Greeff, Anton and Mayer, Marlena and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Impedimetric Biosensor to Enable Fast Evaluation of Gaseous Sterilization Processes}, series = {MDPI Proceedings}, volume = {1}, booktitle = {MDPI Proceedings}, number = {4}, doi = {10.3390/proceedings1040435}, pages = {4 Seiten}, year = {2017}, language = {en} } @inproceedings{MolinnusHardtKaeveretal.2017, author = {Molinnus, Denise and Hardt, Gabriel and K{\"a}ver, Larissa and Willenberg, Holger S. and Poghossian, Arshak and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Detection of Adrenaline Based on Bioelectrocatalytical System to Support Tumor Diagnostic Technology}, series = {MDPI Proceedings}, booktitle = {MDPI Proceedings}, doi = {10.3390/proceedings1040506}, pages = {4 Seiten}, year = {2017}, language = {en} } @article{MuschallikMolinnusBongaertsetal.2017, author = {Muschallik, Lukas and Molinnus, Denise and Bongaerts, Johannes and Pohl, Martina and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Siegert, Petra and Selmer, Thorsten}, title = {(R,R)-Butane-2,3-diol Dehydrogenase from Bacillus clausii DSM 8716T: Cloning and Expression of the bdhA-Gene, and Initial Characterization of Enzyme}, series = {Journal of Biotechnology}, volume = {258}, journal = {Journal of Biotechnology}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0168-1656}, doi = {10.1016/j.jbiotec.2017.07.020}, pages = {41 -- 50}, year = {2017}, abstract = {The gene encoding a putative (R,R)-butane-2,3-diol dehydrogenase (bdhA) from Bacillus clausii DSM 8716T was isolated, sequenced and expressed in Escherichia coli. The amino acid sequence of the encoded protein is only distantly related to previously studied enzymes (identity 33-43\%) and exhibited some uncharted peculiarities. An N-terminally StrepII-tagged enzyme variant was purified and initially characterized. The isolated enzyme catalyzed the (R)-specific oxidation of (R,R)- and meso-butane-2,3-diol to (R)- and (S)-acetoin with specific activities of 12 U/mg and 23 U/mg, respectively. Likewise, racemic acetoin was reduced with a specific activity of up to 115 U/mg yielding a mixture of (R,R)- and meso-butane-2,3-diol, while the enzyme reduced butane-2,3-dione (Vmax 74 U/mg) solely to (R,R)-butane-2,3-diol via (R)-acetoin. For these reactions only activity with the co-substrates NADH/NAD+ was observed. The enzyme accepted a selection of vicinal diketones, α-hydroxy ketones and vicinal diols as alternative substrates. Although the physiological function of the enzyme in B. clausii remains elusive, the data presented herein clearly demonstrates that the encoded enzyme is a genuine (R,R)-butane-2,3-diol dehydrogenase with potential for applications in biocatalysis and sensor development.}, language = {en} } @article{FigueroaMirandaFengShiuetal.2018, author = {Figueroa-Miranda, Gabriela and Feng, Lingyan and Shiu, Simon Chi-Chin and Dirkzwager, Roderick Marshall and Cheung, Yee-Wai and Tanner, Julian Alexander and Sch{\"o}ning, Michael Josef and Offenh{\"a}usser, Andreas and Mayer, Dirk}, title = {Aptamer-based electrochemical biosensor for highly sensitive and selective malaria detection with adjustable dynamic response range and reusability}, series = {Sensor and Actuators B: Chemical}, volume = {255}, journal = {Sensor and Actuators B: Chemical}, number = {P1}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2017.07.117}, pages = {235 -- 243}, year = {2018}, abstract = {Malaria infection remains a significant risk for much of the population of tropical and subtropical areas, particularly in developing countries. Therefore, it is of high importance to develop sensitive, accurate and inexpensive malaria diagnosis tests. Here, we present a novel aptamer-based electrochemical biosensor (aptasensor) for malaria detection by impedance spectroscopy, through the specific recognition between a highly discriminatory DNA aptamer and its target Plasmodium falciparum lactate dehydrogenase (PfLDH). Interestingly, due to the isoelectric point (pI) of PfLDH, the aptasensor response showed an adjustable detection range based on the different protein net-charge at variable pH environments. The specific aptamer recognition allows sensitive protein detection with an expanded detection range and a low detection limit, as well as a high specificity for PfLDH compared to analogous proteins. The specific feasibility of the aptasensor is further demonstrated by detection of the target PfLDH in human serum. Furthermore, the aptasensor can be easily regenerated and thus applied for multiple usages. The robustness, sensitivity, and reusability of the presented aptasensor make it a promising candidate for point-of-care diagnostic systems.}, language = {en} } @article{MoraisGomesSilvaetal.2017, author = {Morais, Paulo V. and Gomes, Vanderley F., Jr. and Silva, Anielle C. A. and Dantas, Noelio O. and Sch{\"o}ning, Michael Josef and Siqueira, Jos{\´e} R., Jr.}, title = {Nanofilm of ZnO nanocrystals/carbon nanotubes as biocompatible layer for enzymatic biosensors in capacitive field-effect devices}, series = {Journal of Materials Science}, volume = {52}, journal = {Journal of Materials Science}, number = {20}, publisher = {Springer}, address = {Berlin}, issn = {1573-4803}, doi = {10.1007/s10853-017-1369-y}, pages = {12314 -- 12325}, year = {2017}, abstract = {The incorporation of nanomaterials that are biocompatible with different types of biological compounds has allowed the development of a new generation of biosensors applied especially in the biomedical field. In particular, the integration of film-based nanomaterials employed in field-effect devices can be interesting to develop biosensors with enhanced properties. In this paper, we studied the fabrication of sensitive nanofilms combining ZnO nanocrystals and carbon nanotubes (CNTs), prepared by means of the layer-by-layer (LbL) technique, in a capacitive electrolyte-insulator-semiconductor (EIS) structure for detecting glucose and urea. The ZnO nanocrystals were incorporated in a polymeric matrix of poly(allylamine) hydrochloride (PAH), and arranged with multi-walled CNTs in a LbL PAH-ZnO/CNTs film architecture onto EIS chips. The electrochemical characterizations were performed by capacitance-voltage and constant capacitance measurements, while the morphology of the films was characterized by atomic force microscopy. The enzymes glucose oxidase and urease were immobilized on film's surface for detection of glucose and urea, respectively. In order to obtain glucose and urea biosensors with optimized amount of sensitive films, we investigated the ideal number of bilayers for each detection system. The glucose biosensor showed better sensitivity and output signal for an LbL PAH-ZnO/CNTs nanofilm with 10 bilayers. On the other hand, the urea biosensor presented enhanced properties even for the first bilayer, exhibiting high sensitivity and output signal. The presence of the LbL PAH-ZnO/CNTs films led to biosensors with better sensitivity and enhanced response signal, demonstrating that the adequate use of nanostructured films is feasible for proof-of-concept biosensors with improved properties that may be employed for biomedical applications.}, language = {en} } @article{HonarvarfardGamellaPoghossianetal.2017, author = {Honarvarfard, Elham and Gamella, Maria and Poghossian, Arshak and Sch{\"o}ning, Michael Josef and Katz, Evgeny}, title = {An enzyme-based reversible Controlled NOT (CNOT) logic gate operating on a semiconductor transducer}, series = {Applied Materials Today}, volume = {9}, journal = {Applied Materials Today}, publisher = {Elsevier}, address = {Amsterdam}, issn = {2352-9407}, doi = {10.1016/j.apmt.2017.08.003}, pages = {266 -- 270}, year = {2017}, abstract = {An enzyme-based biocatalytic system mimicking operation of a logically reversible Controlled NOT (CNOT) gate has been interfaced with semiconductor electronic transducers. Electrolyte-insulator-semiconductor (EIS) structures have been used to transduce chemical changes produced by the enzyme system to an electronically readable capacitive output signal using field-effect features of the EIS device. Two enzymes, urease and esterase, were immobilized on the insulating interface of EIS structure producing local pH changes performing XOR logic operation controlled by various combinations of the input signals represented by urea and ethyl butyrate. Another EIS transducer was functionalized with esterase only, thus performing Identity (ID) logic operation for the ethyl butyrate input. Both semiconductor devices assembled in parallel operated as a logically reversible CNOT gate. The present system, despite its simplicity, demonstrated for the first time logically reversible function of the enzyme system transduced electronically with the semiconductor devices. The biomolecular realization of a CNOT gate interfaced with semiconductors is promising for integration into complex biomolecular networks and future biosensor/biomedical applications.}, language = {en} } @article{RoehlenPilasSchoeningetal.2017, author = {R{\"o}hlen, Desiree and Pilas, Johanna and Sch{\"o}ning, Michael Josef and Selmer, Thorsten}, title = {Development of an amperometric biosensor platform for the combined determination of l-Malic, Fumaric, and l-Aspartic acid}, series = {Applied Biochemistry and Biotechnology}, volume = {183}, journal = {Applied Biochemistry and Biotechnology}, publisher = {Springer}, address = {Berlin}, issn = {1559-0291}, doi = {10.1007/s12010-017-2578-1}, pages = {566 -- 581}, year = {2017}, abstract = {Three amperometric biosensors have been developed for the detection of L-malic acid, fumaric acid, and L -aspartic acid, all based on the combination of a malate-specific dehydrogenase (MDH, EC 1.1.1.37) and diaphorase (DIA, EC 1.8.1.4). The stepwise expansion of the malate platform with the enzymes fumarate hydratase (FH, EC 4.2.1.2) and aspartate ammonia-lyase (ASPA, EC 4.3.1.1) resulted in multi-enzyme reaction cascades and, thus, augmentation of the substrate spectrum of the sensors. Electrochemical measurements were carried out in presence of the cofactor β-nicotinamide adenine dinucleotide (NAD+) and the redox mediator hexacyanoferrate (III) (HCFIII). The amperometric detection is mediated by oxidation of hexacyanoferrate (II) (HCFII) at an applied potential of + 0.3 V vs. Ag/AgCl. For each biosensor, optimum working conditions were defined by adjustment of cofactor concentrations, buffer pH, and immobilization procedure. Under these improved conditions, amperometric responses were linear up to 3.0 mM for L-malate and fumarate, respectively, with a corresponding sensitivity of 0.7 μA mM-1 (L-malate biosensor) and 0.4 μA mM-1 (fumarate biosensor). The L-aspartate detection system displayed a linear range of 1.0-10.0 mM with a sensitivity of 0.09 μA mM-1. The sensor characteristics suggest that the developed platform provides a promising method for the detection and differentiation of the three substrates.}, language = {en} } @article{PilasYaziciSelmeretal.2017, author = {Pilas, Johanna and Yazici, Yasemen and Selmer, Thorsten and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Optimization of an amperometric biosensor array for simultaneous measurement of ethanol, formate, d- and l-lactate}, series = {Electrochimica Acta}, volume = {251}, journal = {Electrochimica Acta}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0013-4686}, doi = {10.1016/j.electacta.2017.07.119}, pages = {256 -- 262}, year = {2017}, abstract = {The immobilization of NAD+-dependent dehydrogenases, in combination with a diaphorase, enables the facile development of multiparametric sensing devices. In this work, an amperometric biosensor array for simultaneous determination of ethanol, formate, d- and l-lactate is presented. Enzyme immobilization on platinum thin-film electrodes was realized by chemical cross-linking with glutaraldehyde. The optimization of the sensor performance was investigated with regard to enzyme loading, glutaraldehyde concentration, pH, cofactor concentration and temperature. Under optimal working conditions (potassium phosphate buffer with pH 7.5, 2.5 mmol L-1 NAD+, 2.0 mmol L-1 ferricyanide, 25 °C and 0.4\% glutaraldehyde) the linear working range and sensitivity of the four sensor elements was improved. Simultaneous and cross-talk free measurements of four different metabolic parameters were performed successfully. The reliable analytical performance of the biosensor array was demonstrated by application in a clarified sample of inoculum sludge. Thereby, a promising approach for on-site monitoring of fermentation processes is provided.}, language = {en} } @article{BreuerMangSchoeningetal.2017, author = {Breuer, Lars and Mang, Thomas and Sch{\"o}ning, Michael Josef and Thoelen, Ronald and Wagner, Torsten}, title = {Investigation of the spatial resolution of a laser-based stimulation process for light-addressable hydrogels with incorporated graphene oxide by means of IR thermography}, series = {Sensors and Actuators A: Physical}, volume = {268}, journal = {Sensors and Actuators A: Physical}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0924-4247}, doi = {10.1016/j.sna.2017.11.031}, pages = {126 -- 132}, year = {2017}, language = {en} } @article{MiyamotoSekiSutoetal.2018, author = {Miyamoto, Koichiro and Seki, Kosuke and Suto, Takeyuki and Werner, Frederik and Wagner, Torsten and Sch{\"o}ning, Michael Josef and Yoshinobu, Tatsuo}, title = {Improved spatial resolution of the chemical imaging sensor with a hybrid illumination that suppresses lateral diffusion of photocarriers}, series = {Sensor and Actuators B: Chemical}, volume = {273}, journal = {Sensor and Actuators B: Chemical}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2018.07.016}, pages = {1328 -- 1333}, year = {2018}, abstract = {The chemical imaging sensor is a semiconductor-based chemical sensor capable of visualizing pH and ion distributions. The spatial resolution depends on the lateral diffusion of photocarriers generated by illumination of the semiconductor substrate. In this study, two types of optical setups, one based on a bundle of optical fibers and the other based on a binocular tube head, were developed to project a hybrid illumination of a modulated light beam and a ring-shaped constant illumination onto the sensor plate. An improved spatial resolution was realized by the ring-shaped constant illumination, which suppressed lateral diffusion of photocarriers by enhanced recombination due to the increased carrier concentration.}, language = {en} } @article{RoehlenPilasDahmenetal.2018, author = {R{\"o}hlen, Desiree and Pilas, Johanna and Dahmen, Markus and Keusgen, Michael and Selmer, Thorsten and Sch{\"o}ning, Michael Josef}, title = {Toward a Hybrid Biosensor System for Analysis of Organic and Volatile Fatty Acids in Fermentation Processes}, series = {Frontiers in Chemistry}, journal = {Frontiers in Chemistry}, number = {6}, publisher = {Frontiers}, address = {Lausanne}, doi = {10.3389/fchem.2018.00284}, pages = {Artikel 284}, year = {2018}, abstract = {Monitoring of organic acids (OA) and volatile fatty acids (VFA) is crucial for the control of anaerobic digestion. In case of unstable process conditions, an accumulation of these intermediates occurs. In the present work, two different enzyme-based biosensor arrays are combined and presented for facile electrochemical determination of several process-relevant analytes. Each biosensor utilizes a platinum sensor chip (14 × 14 mm²) with five individual working electrodes. The OA biosensor enables simultaneous measurement of ethanol, formate, d- and l-lactate, based on a bi-enzymatic detection principle. The second VFA biosensor provides an amperometric platform for quantification of acetate and propionate, mediated by oxidation of hydrogen peroxide. The cross-sensitivity of both biosensors toward potential interferents, typically present in fermentation samples, was investigated. The potential for practical application in complex media was successfully demonstrated in spiked sludge samples collected from three different biogas plants. Thereby, the results obtained by both of the biosensors were in good agreement to the applied reference measurements by photometry and gas chromatography, respectively. The proposed hybrid biosensor system was also used for long-term monitoring of a lab-scale biogas reactor (0.01 m³) for a period of 2 months. In combination with typically monitored parameters, such as gas quality, pH and FOS/TAC (volatile organic acids/total anorganic carbonate), the amperometric measurements of OA and VFA concentration could enhance the understanding of ongoing fermentation processes.}, language = {en} } @article{RodriguesMoraisNordietal.2018, author = {Rodrigues, Raul T. and Morais, Paulo V. and Nordi, Cristina S. F. and Sch{\"o}ning, Michael Josef and Siqueira Jr., Jos{\´e} R. and Caseli, Luciano}, title = {Carbon Nanotubes and Algal Polysaccharides To Enhance the Enzymatic Properties of Urease in Lipid Langmuir-Blodgett Films}, series = {Langmuir}, volume = {34}, journal = {Langmuir}, number = {9}, publisher = {ACS Publications}, address = {Washington, DC}, issn = {1520-5827}, doi = {10.1021/acs.langmuir.7b04317}, pages = {3082 -- 3093}, year = {2018}, abstract = {Algal polysaccharides (extracellular polysaccharides) and carbon nanotubes (CNTs) were adsorbed on dioctadecyldimethylammonium bromide Langmuir monolayers to serve as a matrix for the incorporation of urease. The physicochemical properties of the supramolecular system as a monolayer at the air-water interface were investigated by surface pressure-area isotherms, surface potential-area isotherms, interfacial shear rheology, vibrational spectroscopy, and Brewster angle microscopy. The floating monolayers were transferred to hydrophilic solid supports, quartz, mica, or capacitive electrolyte-insulator-semiconductor (EIS) devices, through the Langmuir-Blodgett (LB) technique, forming mixed films, which were investigated by quartz crystal microbalance, fluorescence spectroscopy, and field emission gun scanning electron microscopy. The enzyme activity was studied with UV-vis spectroscopy, and the feasibility of the thin film as a urea sensor was essayed in an EIS sensor device. The presence of CNT in the enzyme-lipid LB film not only tuned the catalytic activity of urease but also helped to conserve its enzyme activity. Viability as a urease sensor was demonstrated with capacitance-voltage and constant capacitance measurements, exhibiting regular and distinctive output signals over all concentrations used in this work. These results are related to the synergism between the compounds on the active layer, leading to a surface morphology that allowed fast analyte diffusion owing to an adequate molecular accommodation, which also preserved the urease activity. This work demonstrates the feasibility of employing LB films composed of lipids, CNT, algal polysaccharides, and enzymes as EIS devices for biosensing applications.}, language = {en} } @book{SchoeningPoghossian2018, author = {Sch{\"o}ning, Michael Josef and Poghossian, Arshak}, title = {Label-free biosensing: advanced materials, devices and applications}, publisher = {Springer}, address = {Cham}, isbn = {978-3-319-75219-8}, pages = {xii, 480 Seiten ; Illustrationen, Diagramme}, year = {2018}, language = {en} } @incollection{YoshinobuKrauseMiyamotoetal.2018, author = {Yoshinobu, Tatsuo and Krause, Steffi and Miyamoto, Ko-ichiro and Werner, Frederik and Poghossian, Arshak and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {(Bio-)chemical Sensing and Imaging by LAPS and SPIM}, series = {Label-free biosensing: advanced materials, devices and applications}, booktitle = {Label-free biosensing: advanced materials, devices and applications}, publisher = {Springer}, address = {Cham}, isbn = {978-3-319-75219-8}, pages = {103 -- 132}, year = {2018}, abstract = {The light-addressable potentiometric sensor (LAPS) and scanning photo-induced impedance microscopy (SPIM) are two closely related methods to visualise the distributions of chemical species and impedance, respectively, at the interface between the sensing surface and the sample solution. They both have the same field-effect structure based on a semiconductor, which allows spatially resolved and label-free measurement of chemical species and impedance in the form of a photocurrent signal generated by a scanning light beam. In this article, the principles and various operation modes of LAPS and SPIM, functionalisation of the sensing surface for measuring various species, LAPS-based chemical imaging and high-resolution sensors based on silicon-on-sapphire substrates are described and discussed, focusing on their technical details and prospective applications.}, language = {en} } @article{JildehKirchnerBaltesetal.2019, author = {Jildeh, Zaid B. and Kirchner, Patrick and Baltes, Klaus and Wagner, Patrick H. and Sch{\"o}ning, Michael Josef}, title = {Development of an in-line evaporation unit for the production of gas mixtures containing hydrogen peroxide - numerical modeling and experimental results}, series = {International Journal of Heat and Mass Transfer}, volume = {143}, journal = {International Journal of Heat and Mass Transfer}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0017-9310}, doi = {10.1016/j.ijheatmasstransfer.2019.118519}, pages = {Article number 118519}, year = {2019}, abstract = {Hydrogen peroxide (H2O2) is a typical surface sterilization agent for packaging materials used in the pharmaceutical, food and beverage industries. We use the finite-elements method to analyze the conceptual design of an in-line thermal evaporation unit to produce a heated gas mixture of air and evaporated H2O2 solution. For the numerical model, the required phase-transition variables of pure H2O2 solution and of the aerosol mixture are acquired from vapor-liquid equilibrium (VLE) diagrams derived from vapor-pressure formulations. This work combines homogeneous single-phase turbulent flow with heat-transfer physics to describe the operation of the evaporation unit. We introduce the apparent heat-capacity concept to approximate the non-isothermal phase-transition process of the H2O2-containing aerosol. Empirical and analytical functions are defined to represent the temperature- and pressure-dependent material properties of the aqueous H2O2 solution, the aerosol and the gas mixture. To validate the numerical model, the simulation results are compared to experimental data on the heating power required to produce the gas mixture. This shows good agreement with the deviations below 10\%. Experimental observations on the formation of deposits due to the evaporation of stabilized H2O2 solution fits the prediction made from simulation results.}, language = {en} } @article{IkenBronderGoretzkietal.2019, author = {Iken, Heiko and Bronder, Thomas and Goretzki, Alexander and Kriesel, Jana and Ahlborn, Kristina and Gerlach, Frank and Vonau, Winfried and Zander, Willi and Schubert, J{\"u}rgen and Sch{\"o}ning, Michael Josef}, title = {Development of a Combined pH- and Redox-Sensitive Bi-Electrode Glass Thin-Film Sensor}, series = {physica status solidi a : applications and materials sciences}, volume = {216}, journal = {physica status solidi a : applications and materials sciences}, number = {12}, publisher = {Wiley}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201900114}, pages = {1 -- 8}, year = {2019}, language = {en} } @article{OezsoyluKizildagSchoeningetal.2019, author = {{\"O}zsoylu, Dua and Kizildag, Sefa and Sch{\"o}ning, Michael Josef and Wagner, Torsten}, title = {Effect of plasma treatment on the sensor properties of a light-addressable potentiometric sensor (LAPS)}, series = {physica status solidi a : applications and materials sciences}, volume = {216}, journal = {physica status solidi a : applications and materials sciences}, number = {20}, publisher = {Wiley}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201900259}, pages = {8 Seiten}, year = {2019}, abstract = {A light-addressable potentiometric sensor (LAPS) is a field-effect-based (bio-) chemical sensor, in which a desired sensing area on the sensor surface can be defined by illumination. Light addressability can be used to visualize the concentration and spatial distribution of the target molecules, e.g., H+ ions. This unique feature has great potential for the label-free imaging of the metabolic activity of living organisms. The cultivation of those organisms needs specially tailored surface properties of the sensor. O2 plasma treatment is an attractive and promising tool for rapid surface engineering. However, the potential impacts of the technique are carefully investigated for the sensors that suffer from plasma-induced damage. Herein, a LAPS with a Ta2O5 pH-sensitive surface is successfully patterned by plasma treatment, and its effects are investigated by contact angle and scanning LAPS measurements. The plasma duration of 30 s (30 W) is found to be the threshold value, where excessive wettability begins. Furthermore, this treatment approach causes moderate plasma-induced damage, which can be reduced by thermal annealing (10 min at 300 °C). These findings provide a useful guideline to support future studies, where the LAPS surface is desired to be more hydrophilic by O2 plasma treatment.}, language = {en} } @incollection{SchoeningWagnerPoghossianetal.2018, author = {Sch{\"o}ning, Michael Josef and Wagner, Torsten and Poghossian, Arshak and Miyamoto, K.I. and Werner, C.F. and Krause, S. and Yoshinobu, T.}, title = {Light-addressable potentiometric sensors for (bio-)chemical sensing and imaging}, series = {Encyclopedia of Interfacial Chemistry: Surface Science and Electrochemistry. Vol. 7}, booktitle = {Encyclopedia of Interfacial Chemistry: Surface Science and Electrochemistry. Vol. 7}, publisher = {Elsevier}, address = {Amsterdam}, isbn = {9780128097397}, pages = {295 -- 308}, year = {2018}, language = {en} } @article{SchiffelsSelmer2019, author = {Schiffels, Johannes and Selmer, Thorsten}, title = {Combinatorial assembly of ferredoxin-linked modules in Escherichia coli yields a testing platform for Rnf-complexes}, series = {Biotechnology and Bioengineering}, journal = {Biotechnology and Bioengineering}, number = {accepted article}, publisher = {Wiley}, address = {Weinheim}, doi = {10.1002/bit.27079}, pages = {1 -- 36}, year = {2019}, language = {en} } @article{DantismRoehlenSelmeretal.2019, author = {Dantism, Shahriar and R{\"o}hlen, Desiree and Selmer, Thorsten and Wagner, Torsten and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Quantitative differential monitoring of the metabolic activity of Corynebacterium glutamicum cultures utilizing a light-addressable potentiometric sensor system}, series = {Biosensors and Bioelectronics}, volume = {139}, journal = {Biosensors and Bioelectronics}, publisher = {Elsevier}, address = {Amsterdam}, doi = {10.1016/j.bios.2019.111332}, pages = {Artikel 111332}, year = {2019}, language = {en} } @article{OezsoyluKizildagSchoeningetal.2020, author = {{\"O}zsoylu, Dua and Kizildag, Sefa and Sch{\"o}ning, Michael Josef and Wagner, Torsten}, title = {Differential chemical imaging of extracellular acidification within microfluidic channels using a plasma-functionalized light-addressable potentiometric sensor (LAPS)}, series = {Physics in Medicine}, volume = {10}, journal = {Physics in Medicine}, number = {100030}, publisher = {Elsevier}, address = {Amsterdam}, issn = {2352-4510}, doi = {10.1016/j.phmed.2020.100030}, pages = {8}, year = {2020}, abstract = {Extracellular acidification is a basic indicator for alterations in two vital metabolic pathways: glycolysis and cellular respiration. Measuring these alterations by monitoring extracellular acidification using cell-based biosensors such as LAPS plays an important role in studying these pathways whose disorders are associated with numerous diseases including cancer. However, the surface of the biosensors must be specially tailored to ensure high cell compatibility so that cells can represent more in vivo-like behavior, which is critical to gain more realistic in vitro results from the analyses, e.g., drug discovery experiments. In this work, O2 plasma patterning on the LAPS surface is studied to enhance surface features of the sensor chip, e.g., wettability and biofunctionality. The surface treated with O2 plasma for 30 s exhibits enhanced cytocompatibility for adherent CHO-K1 cells, which promotes cell spreading and proliferation. The plasma-modified LAPS chip is then integrated into a microfluidic system, which provides two identical channels to facilitate differential measurements of the extracellular acidification of CHO-K1 cells. To the best of our knowledge, it is the first time that extracellular acidification within microfluidic channels is quantitatively visualized as differential (bio-)chemical images.}, language = {en} } @article{MuschallikKippReckeretal.2020, author = {Muschallik, Lukas and Kipp, Carina Ronja and Recker, Inga and Bongaerts, Johannes and Pohl, Martina and Gelissen, Melanie and Sch{\"o}ning, Michael Josef and Selmer, Thorsten and Siegert, Petra}, title = {Synthesis of α-hydroxy ketones and vicinal diols with the Bacillus licheniformis DSM 13T butane-2, 3-diol dehydrogenase}, series = {Journal of Biotechnology}, volume = {202}, journal = {Journal of Biotechnology}, number = {Vol. 324}, publisher = {Elsevier}, address = {Amsterdam}, isbn = {2590-1559}, doi = {10.1016/j.jbiotec.2020.09.016}, pages = {61 -- 70}, year = {2020}, abstract = {The enantioselective synthesis of α-hydroxy ketones and vicinal diols is an intriguing field because of the broad applicability of these molecules. Although, butandiol dehydrogenases are known to play a key role in the production of 2,3-butandiol, their potential as biocatalysts is still not well studied. Here, we investigate the biocatalytic properties of the meso-butanediol dehydrogenase from Bacillus licheniformis DSM 13T (BlBDH). The encoding gene was cloned with an N-terminal StrepII-tag and recombinantly overexpressed in E. coli. BlBDH is highly active towards several non-physiological diketones and α-hydroxyketones with varying aliphatic chain lengths or even containing phenyl moieties. By adjusting the reaction parameters in biotransformations the formation of either the α-hydroxyketone intermediate or the diol can be controlled.}, language = {en} } @article{MolinnusIkenJohnenetal.2022, author = {Molinnus, Denise and Iken, Heiko and Johnen, Anna Lynn and Richstein, Benjamin and Hellmich, Lena and Poghossian, Arshak and Knoch, Joachim and Sch{\"o}ning, Michael Josef}, title = {Miniaturized pH-Sensitive Field-Effect Capacitors with Ultrathin Ta₂O₅ Films Prepared by Atomic Layer Deposition}, series = {physica status solidi (a) applications and materials science}, volume = {219}, journal = {physica status solidi (a) applications and materials science}, number = {8}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.202100660}, pages = {7 Seiten}, year = {2022}, abstract = {Miniaturized electrolyte-insulator-semiconductor capacitors (EISCAPs) with ultrathin gate insulators have been studied in terms of their pH-sensitive sensor characteristics: three different EISCAP systems consisting of Al-p-Si-Ta2O5(5 nm), Al-p-Si-Si3N4(1 or 2 nm)-Ta2O5 (5 nm), and Al-p-Si-SiO2(3.6 nm)-Ta2O5(5 nm) layer structures are characterized in buffer solution with different pH values by means of capacitance-voltage and constant capacitance method. The SiO2 and Si3N4 gate insulators are deposited by rapid thermal oxidation and rapid thermal nitridation, respectively, whereas the Ta2O5 film is prepared by atomic layer deposition. All EISCAP systems have a clear pH response, favoring the stacked gate insulators SiO2-Ta2O5 when considering the overall sensor characteristics, while the Si3N4(1 nm)-Ta2O5 stack delivers the largest accumulation capacitance (due to the lower equivalent oxide thickness) and a higher steepness in the slope of the capacitance-voltage curve among the studied stacked gate insulator systems.}, language = {en} } @article{OliveiraMolinnusBegingetal.2021, author = {Oliveira, Danilo A. and Molinnus, Denise and Beging, Stefan and Siqueira Jr, Jos{\´e} R. and Sch{\"o}ning, Michael Josef}, title = {Biosensor Based on Self-Assembled Films of Graphene Oxide and Polyaniline Using a Field-Effect Device Platform}, series = {physica status solidi (a) applications and materials science}, volume = {218}, journal = {physica status solidi (a) applications and materials science}, number = {13}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.202000747}, pages = {1 -- 9}, year = {2021}, abstract = {A new functionalization method to modify capacitive electrolyte-insulator-semiconductor (EIS) structures with nanofilms is presented. Layers of polyallylamine hydrochloride (PAH) and graphene oxide (GO) with the compound polyaniline:poly(2-acrylamido-2-methyl-1-propanesulfonic acid) (PANI:PAAMPSA) are deposited onto a p-Si/SiO2 chip using the layer-by-layer technique (LbL). Two different enzymes (urease and penicillinase) are separately immobilized on top of a five-bilayer stack of the PAH:GO/PANI:PAAMPSA-modified EIS chip, forming a biosensor for detection of urea and penicillin, respectively. Electrochemical characterization is performed by constant capacitance (ConCap) measurements, and the film morphology is characterized by atomic force microscopy (AFM) and scanning electron microscopy (SEM). An increase in the average sensitivity of the modified biosensors (EIS-nanofilm-enzyme) of around 15\% is found in relation to sensors, only carrying the enzyme but without the nanofilm (EIS-enzyme). In this sense, the nanofilm acts as a stable bioreceptor onto the EIS chip improving the output signal in terms of sensitivity and stability.}, language = {en} } @article{WeldenNagamineKomesuWagneretal.2021, author = {Welden, Rene and Nagamine Komesu, Cindy A. and Wagner, Patrick H. and Sch{\"o}ning, Michael Josef and Wagner, Torsten}, title = {Photoelectrochemical enzymatic penicillin biosensor: A proof-of-concept experiment}, series = {Electrochemical Science Advances}, volume = {2}, journal = {Electrochemical Science Advances}, number = {4}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {2698-5977}, doi = {10.1002/elsa.202100131}, pages = {1 -- 5}, year = {2021}, abstract = {Photoelectrochemical (PEC) biosensors are a rather novel type of biosensors thatutilizelighttoprovideinformationaboutthecompositionofananalyte,enablinglight-controlled multi-analyte measurements. For enzymatic PEC biosensors,amperometric detection principles are already known in the literature. In con-trast, there is only a little information on H+-ion sensitive PEC biosensors. Inthis work, we demonstrate the detection of H+ions emerged by H+-generatingenzymes, exemplarily demonstrated with penicillinase as a model enzyme on atitanium dioxide photoanode. First, we describe the pH sensitivity of the sensorand study possible photoelectrocatalytic reactions with penicillin. Second, weshow the enzymatic PEC detection of penicillin.}, language = {en} } @article{JablonskiPoghossianKeusgenetal.2021, author = {Jablonski, Melanie and Poghossian, Arshak and Keusgen, Michael and Wege, Christina and Sch{\"o}ning, Michael Josef}, title = {Detection of plant virus particles with a capacitive field-effect sensor}, series = {Analytical and Bioanalytical Chemistry}, volume = {413}, journal = {Analytical and Bioanalytical Chemistry}, publisher = {Springer Nature}, address = {Cham}, issn = {1618-2650}, doi = {10.1007/s00216-021-03448-8}, pages = {5669 -- 5678}, year = {2021}, abstract = {Plant viruses are major contributors to crop losses and induce high economic costs worldwide. For reliable, on-site and early detection of plant viral diseases, portable biosensors are of great interest. In this study, a field-effect SiO2-gate electrolyte-insulator-semiconductor (EIS) sensor was utilized for the label-free electrostatic detection of tobacco mosaic virus (TMV) particles as a model plant pathogen. The capacitive EIS sensor has been characterized regarding its TMV sensitivity by means of constant-capacitance method. The EIS sensor was able to detect biotinylated TMV particles from a solution with a TMV concentration as low as 0.025 nM. A good correlation between the registered EIS sensor signal and the density of adsorbed TMV particles assessed from scanning electron microscopy images of the SiO2-gate chip surface was observed. Additionally, the isoelectric point of the biotinylated TMV particles was determined via zeta potential measurements and the influence of ionic strength of the measurement solution on the TMV-modified EIS sensor signal has been studied.}, language = {en} } @article{PoghossianWernerBuniatyanetal.2017, author = {Poghossian, Arshak and Werner, Frederik and Buniatyan, V. V. and Wagner, Torsten and Miamoto, K. and Yoshinobu, T. and Sch{\"o}ning, Michael Josef}, title = {Towards addressability of light-addressable potentiometric sensors: Shunting effect of non-illuminated region and cross-talk}, series = {Sensor and Actuators B: Chemical}, journal = {Sensor and Actuators B: Chemical}, number = {244}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2017.01.047}, pages = {1071 -- 1079}, year = {2017}, abstract = {The LAPS (light-addressable potentiometric sensor) platform is one of the most attractive approaches for chemical and biological sensing with many applications ranging from pH and ion/analyte concentration measurements up to cell metabolism detection and chemical imaging. However, although it is generally accepted that LAPS measurements are spatially resolved, the light-addressability feature of LAPS devices has not been discussed in detail so far. In this work, an extended electrical equivalent-circuit model of the LAPS has been presented, which takes into account possible cross-talk effects due to the capacitive coupling of the non-illuminated region. A shunting effect of the non-illuminated area on the measured photocurrent and addressability of LAPS devices has been studied. It has been shown, that the measured photocurrent will be determined not only by the local interfacial potential in the illuminated region but also by possible interfacial potential changes in the non-illuminated region, yielding cross-talk effects. These findings were supported by the experimental investigations of a penicillin-sensitive multi-spot LAPS and a metal-insulator-semiconductor LAPS as model systems.}, language = {en} } @inproceedings{BaeckerKochGeigeretal.2016, author = {B{\"a}cker, Matthias and Koch, C. and Geiger, F. and Eber, F. and Gliemann, H. and Poghossian, Arshak and Sch{\"o}ning, Michael Josef}, title = {A New Class of Biosensors Based on Tobacco Mosaic Virus and Coat Proteins as Enzyme Nanocarrier}, series = {Procedia Engineering}, volume = {Vol. 168}, booktitle = {Procedia Engineering}, issn = {1877-7058}, doi = {10.1016/j.proeng.2016.11.228}, pages = {618 -- 621}, year = {2016}, language = {en} } @article{KatzPoghossianSchoening2017, author = {Katz, Evgeny and Poghossian, Arshak and Sch{\"o}ning, Michael Josef}, title = {Enzyme-based logic gates and circuits - analytical applications and interfacing with electronics}, series = {Analytical and Bioanalytical Chemistry}, volume = {409}, journal = {Analytical and Bioanalytical Chemistry}, publisher = {Springer}, address = {Berlin}, issn = {1618-2650}, doi = {10.1007/s00216-016-0079-7}, pages = {81 -- 94}, year = {2017}, abstract = {The paper is an overview of enzyme-based logic gates and their short circuits, with specific examples of Boolean AND and OR gates, and concatenated logic gates composed of multi-step enzyme-biocatalyzed reactions. Noise formation in the biocatalytic reactions and its decrease by adding a "filter" system, converting convex to sigmoid response function, are discussed. Despite the fact that the enzyme-based logic gates are primarily considered as components of future biomolecular computing systems, their biosensing applications are promising for immediate practical use. Analytical use of the enzyme logic systems in biomedical and forensic applications is discussed and exemplified with the logic analysis of biomarkers of various injuries, e.g., liver injury, and with analysis of biomarkers characteristic of different ethnicity found in blood samples on a crime scene. Interfacing of enzyme logic systems with modified electrodes and semiconductor devices is discussed, giving particular attention to the interfaces functionalized with signal-responsive materials. Future perspectives in the design of the biomolecular logic systems and their applications are discussed in the conclusion.}, language = {en} } @inproceedings{PoghossianBronderSchejaetal.2016, author = {Poghossian, Arshak and Bronder, Thomas and Scheja, S. and Wu, Chunsheng and Metzger-Boddien, C. and Keusgen, M. and Sch{\"o}ning, Michael Josef}, title = {Label-free Electrostatic Detection of DNA Amplification by PCR Using Capacitive Field-effect Devices}, series = {Procedia Engineering}, volume = {Vol. 168}, booktitle = {Procedia Engineering}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1877-7058}, doi = {10.1016/j.proeng.2016.11.512}, pages = {514 -- 517}, year = {2016}, abstract = {A capacitive field-effect EIS (electrolyte-insulator-semiconductor) sensor modified with a positively charged weak polyelectrolyte of poly(allylamine hydrochloride) (PAH)/single-stranded probe DNA (ssDNA) bilayer has been used for a label-free electrostatic detection of pathogen-specific DNA amplification via polymerase chain reaction (PCR). The sensor is able to distinguish between positive and negative PCR solutions, to detect the existence of target DNA amplicons in PCR samples and thus, can be used as tool for a quick verification of DNA amplification and the successful PCR process.}, language = {en} } @article{HonarvarfardGamellaChannaveerappaetal.2017, author = {Honarvarfard, Elham and Gamella, Maria and Channaveerappa, Devika and Darie, Costel C. and Poghossian, Arshak and Sch{\"o}ning, Michael Josef and Katz, Evgeny}, title = {Electrochemically Stimulated Insulin Release from a Modified Graphene-functionalized Carbon Fiber Electrode}, series = {Electroanalysis}, volume = {29}, journal = {Electroanalysis}, number = {6}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1521-4109}, doi = {10.1002/elan.201700095}, pages = {1543 -- 1553}, year = {2017}, abstract = {A graphene-functionalized carbon fiber electrode was modified with adsorbed polyethylenimine to introduce amino functionalities and then with trigonelline and 4-carboxyphenylboronic acid covalently bound to the amino groups. The trigonelline species containing quarterized pyridine groups produced positive charge on the electrode surface regardless of the pH value, while the phenylboronic acid species were neutral below pH 8 and negatively charged above pH 9 (note that their pKa=8.4). The total charge on the monolayer-modified electrode was positive at the neutral pH and negative at pH > 9. Note that 4-carboxyphenylboronic acid was attached to the electrode surface in molar excess to trigonelline, thus allowing the negative charge to dominate on the electrode surface at basic pH. Negatively charged fluorescent dye-labeled insulin (insulin-FITC) was loaded on the modified electrode surface at pH 7.0 due to its electrostatic attraction to the positively charged interface. The local pH in close vicinity to the electrode surface was increased to ca. 9-10 due to consumption of H+ ions upon electrochemical reduction of oxygen proceeding at the potential of -1.0 V (vs. Ag/AgCl) applied on the modified electrode. The process resulted in recharging of the electrode surface to the negative value due to the formation of the negative charge on the phenylboronic acid groups, thus resulting in the electrostatic repulsion of insulin-FITC and stimulating its release from the electrode surface. The insulin release was characterized by fluorescence spectroscopy (using the FITC-labeled insulin), by electrochemical measurements on an iridium oxide, IrOx, electrode and by mass spectrometry. The graphene-functionalized carbon fiber electrode demonstrated significant advantages in the signal-stimulated insulin release comparing with the carbon fiber electrode without the graphene species.}, language = {en} } @article{JildehWagnerSchoening2021, author = {Jildeh, Zaid B. and Wagner, Patrick H. and Sch{\"o}ning, Michael Josef}, title = {Sterilization of Objects, Products, and Packaging Surfaces and Their Characterization in Different Fields of Industry: The Status in 2020}, series = {physica status solidi (a) applications and materials science}, volume = {218}, journal = {physica status solidi (a) applications and materials science}, number = {13}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.202000732}, pages = {27 Seiten}, year = {2021}, abstract = {The treatment method to deactivate viable microorganisms from objects or products is termed sterilization. There are multiple forms of sterilization, each intended to be applied for a specific target, which depends on—but not limited to—the thermal, physical, and chemical stability of that target. Herein, an overview on the currently used sterilization processes in the global market is provided. Different sterilization techniques are grouped under a category that describes the method of treatment: radiation (gamma, electron beam, X-ray, and ultraviolet), thermal (dry and moist heat), and chemical (ethylene oxide, ozone, chlorine dioxide, and hydrogen peroxide). For each sterilization process, the typical process parameters as defined by regulations and the mode of antimicrobial activity are summarized. Finally, the recommended microorganisms that are used as biological indicators to validate sterilization processes in accordance with the rules that are established by various regulatory agencies are summarized.}, language = {en} } @article{MolinnusDrinicIkenetal.2021, author = {Molinnus, Denise and Drinic, Aleksander and Iken, Heiko and Kr{\"o}ger, Nadja and Zinser, Max and Smeets, Ralf and K{\"o}pf, Marius and Kopp, Alexander and Sch{\"o}ning, Michael Josef}, title = {Towards a flexible electrochemical biosensor fabricated from biocompatible Bombyx mori silk}, series = {Biosensors and Bioelectronics}, volume = {183}, journal = {Biosensors and Bioelectronics}, number = {Art. 113204}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0956-5663}, doi = {10.1016/j.bios.2021.113204}, year = {2021}, language = {en} } @article{YoshinobuSchoening2021, author = {Yoshinobu, Tatsuo and Sch{\"o}ning, Michael Josef}, title = {Light-addressable potentiometric sensors (LAPS) for cell monitoring and biosensing}, series = {Current Opinion in Electrochemistry}, journal = {Current Opinion in Electrochemistry}, number = {In Press, Journal Pre-proof}, publisher = {Elsevier}, address = {Amsterdam}, issn = {2451-9103}, doi = {10.1016/j.coelec.2021.100727}, year = {2021}, language = {en} } @phdthesis{Bronder2020, author = {Bronder, Thomas}, title = {Label-free detection of tuberculosis DNA with capacitive field-effect biosensors}, publisher = {Philipps-Universit{\"a}t Marburg}, address = {Marburg}, doi = {10.17192/z2021.0056}, pages = {X, 162 S}, year = {2020}, language = {en} } @article{JablonskiMuenstermannNorketal.2021, author = {Jablonski, Melanie and M{\"u}nstermann, Felix and Nork, Jasmina and Molinnus, Denise and Muschallik, Lukas and Bongaerts, Johannes and Wagner, Torsten and Keusgen, Michael and Siegert, Petra and Sch{\"o}ning, Michael Josef}, title = {Capacitive field-effect biosensor applied for the detection of acetoin in alcoholic beverages and fermentation broths}, series = {physica status solidi (a) applications and materials science}, volume = {218}, journal = {physica status solidi (a) applications and materials science}, number = {13}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.202000765}, pages = {7 Seiten}, year = {2021}, abstract = {An acetoin biosensor based on a capacitive electrolyte-insulator-semiconductor (EIS) structure modified with the enzyme acetoin reductase, also known as butane-2,3-diol dehydrogenase (Bacillus clausii DSM 8716ᵀ), is applied for acetoin detection in beer, red wine, and fermentation broth samples for the first time. The EIS sensor consists of an Al/p-Si/SiO₂/Ta₂O₅ layer structure with immobilized acetoin reductase on top of the Ta₂O₅ transducer layer by means of crosslinking via glutaraldehyde. The unmodified and enzyme-modified sensors are electrochemically characterized by means of leakage current, capacitance-voltage, and constant capacitance methods, respectively.}, language = {en} } @article{WertIkenSchoeningetal.2021, author = {Wert, Stefan and Iken, Heiko and Sch{\"o}ning, Michael Josef and Matysik, Frank-Michael}, title = {Development of a temperature-pulse enhanced electrochemical glucose biosensor and characterization of its stability via scanning electrochemical microscopy}, series = {Electroanalysis}, journal = {Electroanalysis}, number = {Early View}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1521-4109}, doi = {10.1002/elan.202100089}, year = {2021}, abstract = {Glucose oxidase (GOx) is an enzyme frequently used in glucose biosensors. As increased temperatures can enhance the performance of electrochemical sensors, we investigated the impact of temperature pulses on GOx that was drop-coated on flattened Pt microwires. The wires were heated by an alternating current. The sensitivity towards glucose and the temperature stability of GOx was investigated by amperometry. An up to 22-fold increase of sensitivity was observed. Spatially resolved enzyme activity changes were investigated via scanning electrochemical microscopy. The application of short (<100 ms) heat pulses was associated with less thermal inactivation of the immobilized GOx than long-term heating.}, language = {en} } @article{JablonskiPoghossianSeverinetal.2021, author = {Jablonski, Melanie and Poghossian, Arshak and Severin, Robin and Keusgen, Michael and Wege, Christian and Sch{\"o}ning, Michael Josef}, title = {Capacitive Field-Effect Biosensor Studying Adsorption of Tobacco Mosaic Virus Particles}, series = {Micromachines}, volume = {12}, journal = {Micromachines}, number = {1}, publisher = {MDPI}, address = {Basel}, doi = {10.3390/mi12010057}, pages = {Artikel 57}, year = {2021}, abstract = {Plant virus-like particles, and in particular, tobacco mosaic virus (TMV) particles, are increasingly being used in nano- and biotechnology as well as for biochemical sensing purposes as nanoscaffolds for the high-density immobilization of receptor molecules. The sensitive parameters of TMV-assisted biosensors depend, among others, on the density of adsorbed TMV particles on the sensor surface, which is affected by both the adsorption conditions and surface properties of the sensor. In this work, Ta₂O₅-gate field-effect capacitive sensors have been applied for the label-free electrical detection of TMV adsorption. The impact of the TMV concentration on both the sensor signal and the density of TMV particles adsorbed onto the Ta₂O₅-gate surface has been studied systematically by means of field-effect and scanning electron microscopy methods. In addition, the surface density of TMV particles loaded under different incubation times has been investigated. Finally, the field-effect sensor also demonstrates the label-free detection of penicillinase immobilization as model bioreceptor on TMV particles.}, language = {en} } @article{GivanoudiCornelisRasschaertetal.2021, author = {Givanoudi, Stella and Cornelis, Peter and Rasschaert, Geertrui and Wackers, Gideon and Iken, Heiko and Rolka, David and Yongabi, Derick and Robbens, Johan and Sch{\"o}ning, Michael Josef and Heyndrickx, Marc and Wagner, Patrick}, title = {Selective Campylobacter detection and quantification in poultry: A sensor tool for detecting the cause of a common zoonosis at its source}, series = {Sensors and Actuators B: Chemical}, journal = {Sensors and Actuators B: Chemical}, number = {In Press, Journal Pre-proof}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2021.129484}, pages = {Article 129484}, year = {2021}, language = {en} } @article{VahidpourGuthmanArreolaetal.2022, author = {Vahidpour, Farnoosh and Guthman, Eric and Arreola, Julia and Alghazali, Yousef H. M. and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Assessment of Various Process Parameters for Optimized Sterilization Conditions Using a Multi-Sensing Platform}, series = {Foods}, volume = {11}, journal = {Foods}, number = {5}, publisher = {MDPI}, address = {Basel}, issn = {2304-8158}, doi = {10.3390/foods11050660}, pages = {Artikel 660}, year = {2022}, abstract = {In this study, an online multi-sensing platform was engineered to simultaneously evaluate various process parameters of food package sterilization using gaseous hydrogen peroxide (H₂O₂). The platform enabled the validation of critical aseptic parameters. In parallel, one series of microbiological count reduction tests was performed using highly resistant spores of B. atrophaeus DSM 675 to act as the reference method for sterility validation. By means of the multi-sensing platform together with microbiological tests, we examined sterilization process parameters to define the most effective conditions with regards to the highest spore kill rate necessary for aseptic packaging. As these parameters are mutually associated, a correlation between different factors was elaborated. The resulting correlation indicated the need for specific conditions regarding the applied H₂O₂ gas temperature, the gas flow and concentration, the relative humidity and the exposure time. Finally, the novel multi-sensing platform together with the mobile electronic readout setup allowed for the online and on-site monitoring of the sterilization process, selecting the best conditions for sterility and, at the same time, reducing the use of the time-consuming and costly microbiological tests that are currently used in the food package industry.}, language = {en} } @article{PoghossianWeldenBuniatyanetal.2021, author = {Poghossian, Arshak and Welden, Rene and Buniatyan, Vahe V. and Sch{\"o}ning, Michael Josef}, title = {An Array of On-Chip Integrated, Individually Addressable Capacitive Field-Effect Sensors with Control Gate: Design and Modelling}, series = {Sensors}, volume = {21}, journal = {Sensors}, number = {18}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s21186161}, pages = {17}, year = {2021}, abstract = {The on-chip integration of multiple biochemical sensors based on field-effect electrolyte-insulator-semiconductor capacitors (EISCAP) is challenging due to technological difficulties in realization of electrically isolated EISCAPs on the same Si chip. In this work, we present a new simple design for an array of on-chip integrated, individually electrically addressable EISCAPs with an additional control gate (CG-EISCAP). The existence of the CG enables an addressable activation or deactivation of on-chip integrated individual CG-EISCAPs by simple electrical switching the CG of each sensor in various setups, and makes the new design capable for multianalyte detection without cross-talk effects between the sensors in the array. The new designed CG-EISCAP chip was modelled in so-called floating/short-circuited and floating/capacitively-coupled setups, and the corresponding electrical equivalent circuits were developed. In addition, the capacitance-voltage curves of the CG-EISCAP chip in different setups were simulated and compared with that of a single EISCAP sensor. Moreover, the sensitivity of the CG-EISCAP chip to surface potential changes induced by biochemical reactions was simulated and an impact of different parameters, such as gate voltage, insulator thickness and doping concentration in Si, on the sensitivity has been discussed.}, language = {en} } @article{PourshahidiAchtsnichtNambipareecheeetal.2021, author = {Pourshahidi, Ali Mohammad and Achtsnicht, Stefan and Nambipareechee, Mrinal Murali and Offenh{\"a}usser, Andreas and Krause, Hans-Joachim}, title = {Multiplex detection of magnetic beads using offset field dependent frequency mixing magnetic detection}, series = {Sensors}, volume = {21}, journal = {Sensors}, number = {17}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s21175859}, pages = {16 Seiten}, year = {2021}, abstract = {Magnetic immunoassays employing Frequency Mixing Magnetic Detection (FMMD) have recently become increasingly popular for quantitative detection of various analytes. Simultaneous analysis of a sample for two or more targets is desirable in order to reduce the sample amount, save consumables, and save time. We show that different types of magnetic beads can be distinguished according to their frequency mixing response to a two-frequency magnetic excitation at different static magnetic offset fields. We recorded the offset field dependent FMMD response of two different particle types at frequencies ƒ₁ + n⋅ƒ₂, n = 1, 2, 3, 4 with ƒ₁ = 30.8 kHz and ƒ₂ = 63 Hz. Their signals were clearly distinguishable by the locations of the extremes and zeros of their responses. Binary mixtures of the two particle types were prepared with different mixing ratios. The mixture samples were analyzed by determining the best linear combination of the two pure constituents that best resembled the measured signals of the mixtures. Using a quadratic programming algorithm, the mixing ratios could be determined with an accuracy of greater than 14\%. If each particle type is functionalized with a different antibody, multiplex detection of two different analytes becomes feasible.}, language = {en} } @article{HaegerGrankinWagner2023, author = {Haeger, Gerrit and Grankin, Alina and Wagner, Michaela}, title = {Construction of an Aspergillus oryzae triple amylase deletion mutant as a chassis to evaluate industrially relevant amylases using multiplex CRISPR/Cas9 editing technology}, series = {Applied Research}, journal = {Applied Research}, number = {Early View}, publisher = {Wiley-VCH}, issn = {2702-4288}, doi = {10.1002/appl.202200106}, pages = {1 -- 15}, year = {2023}, abstract = {Aspergillus oryzae is an industrially relevant organism for the secretory production of heterologous enzymes, especially amylases. The activities of potential heterologous amylases, however, cannot be quantified directly from the supernatant due to the high background activity of native α-amylase. This activity is caused by the gene products of amyA, amyB, and amyC. In this study, an in vitro CRISPR/Cas9 system was established in A. oryzae to delete these genes simultaneously. First, pyrG of A. oryzae NSAR1 was mutated by exploiting NHEJ to generate a counter-selection marker. Next, all amylase genes were deleted simultaneously by co-transforming a repair template carrying pyrG of Aspergillus nidulans and flanking sequences of amylase gene loci. The rate of obtained triple knock-outs was 47\%. We showed that triple knockouts do not retain any amylase activity in the supernatant. The established in vitro CRISPR/Cas9 system was used to achieve sequence-specific knock-in of target genes. The system was intended to incorporate a single copy of the gene of interest into the desired host for the development of screening methods. Therefore, an integration cassette for the heterologous Fpi amylase was designed to specifically target the amyB locus. The site-specific integration rate of the plasmid was 78\%, with exceptional additional integrations. Integration frequency was assessed via qPCR and directly correlated with heterologous amylase activity. Hence, we could compare the efficiency between two different signal peptides. In summary, we present a strategy to exploit CRISPR/Cas9 for gene mutation, multiplex knock-out, and the targeted knock-in of an expression cassette in A. oryzae. Our system provides straightforward strain engineering and paves the way for development of fungal screening systems.}, language = {en} } @article{VahidpourAlghazaliAkcaetal.2022, author = {Vahidpour, Farnoosh and Alghazali, Yousef and Akca, Sevilay and Hommes, Gregor and Sch{\"o}ning, Michael Josef}, title = {An Enzyme-Based Interdigitated Electrode-Type Biosensor for Detecting Low Concentrations of H₂O₂ Vapor/Aerosol}, series = {Chemosensors}, volume = {10}, journal = {Chemosensors}, number = {6}, publisher = {MDPI}, address = {Basel}, issn = {2227-9040}, doi = {10.3390/chemosensors10060202}, pages = {Arikel 202}, year = {2022}, abstract = {This work introduces a novel method for the detection of H₂O₂ vapor/aerosol of low concentrations, which is mainly applied in the sterilization of equipment in medical industry. Interdigitated electrode (IDE) structures have been fabricated by means of microfabrication techniques. A differential setup of IDEs was prepared, containing an active sensor element (active IDE) and a passive sensor element (passive IDE), where the former was immobilized with an enzymatic membrane of horseradish peroxidase that is selective towards H₂O₂. Changes in the IDEs' capacitance values (active sensor element versus passive sensor element) under H₂O₂ vapor/aerosol atmosphere proved the detection in the concentration range up to 630 ppm with a fast response time (<60 s). The influence of relative humidity was also tested with regard to the sensor signal, showing no cross-sensitivity. The repeatability assessment of the IDE biosensors confirmed their stable capacitive signal in eight subsequent cycles of exposure to H₂O₂ vapor/aerosol. Room-temperature detection of H₂O₂ vapor/aerosol with such miniaturized biosensors will allow a future three-dimensional, flexible mapping of aseptic chambers and help to evaluate sterilization assurance in medical industry.}, language = {en} } @article{PoghossianKarschuckWagneretal.2022, author = {Poghossian, Arshak and Karschuck, Tobias and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Field-Effect Capacitors Decorated with Ligand-Stabilized Gold Nanoparticles: Modeling and Experiments}, series = {Biosensors}, volume = {12}, journal = {Biosensors}, number = {5}, publisher = {MDPI}, address = {Basel}, issn = {2079-6374}, doi = {10.3390/bios12050334}, pages = {Artikel 334}, year = {2022}, abstract = {Nanoparticles are recognized as highly attractive tunable materials for designing field-effect biosensors with enhanced performance. In this work, we present a theoretical model for electrolyte-insulator-semiconductor capacitors (EISCAP) decorated with ligand-stabilized charged gold nanoparticles. The charged AuNPs are taken into account as additional, nanometer-sized local gates. The capacitance-voltage (C-V) curves and constant-capacitance (ConCap) signals of the AuNP-decorated EISCAPs have been simulated. The impact of the AuNP coverage on the shift of the C-V curves and the ConCap signals was also studied experimentally on Al-p-Si-SiO₂ EISCAPs decorated with positively charged aminooctanethiol-capped AuNPs. In addition, the surface of the EISCAPs, modified with AuNPs, was characterized by scanning electron microscopy for different immobilization times of the nanoparticles.}, language = {en} } @article{PourshahidiAchtsnichtOffenhaeusseretal.2022, author = {Pourshahidi, Ali Mohammad and Achtsnicht, Stefan and Offenh{\"a}usser, Andreas and Krause, Hans-Joachim}, title = {Frequency Mixing Magnetic Detection Setup Employing Permanent Ring Magnets as a Static Offset Field Source}, series = {Sensors}, volume = {22}, journal = {Sensors}, number = {22}, editor = {Offenh{\"a}usser, Andreas}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s22228776}, pages = {12 Seiten}, year = {2022}, abstract = {Frequency mixing magnetic detection (FMMD) has been explored for its applications in fields of magnetic biosensing, multiplex detection of magnetic nanoparticles (MNP) and the determination of core size distribution of MNP samples. Such applications rely on the application of a static offset magnetic field, which is generated traditionally with an electromagnet. Such a setup requires a current source, as well as passive or active cooling strategies, which directly sets a limitation based on the portability aspect that is desired for point of care (POC) monitoring applications. In this work, a measurement head is introduced that involves the utilization of two ring-shaped permanent magnets to generate a static offset magnetic field. A steel cylinder in the ring bores homogenizes the field. By variation of the distance between the ring magnets and of the thickness of the steel cylinder, the magnitude of the magnetic field at the sample position can be adjusted. Furthermore, the measurement setup is compared to the electromagnet offset module based on measured signals and temperature behavior.}, language = {en} } @article{KarschuckKaulenPoghossianetal.2021, author = {Karschuck, Tobias and Kaulen, Corinna and Poghossian, Arshak and Wagner, Patrick H. and Sch{\"o}ning, Michael Josef}, title = {Gold nanoparticle-modified capacitive field-effect sensors: Studying the surface density of nanoparticles and coupling of charged polyelectrolyte macromolecules}, series = {Electrochemical Science Advances}, volume = {2}, journal = {Electrochemical Science Advances}, number = {5}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {0938-5193}, doi = {10.1002/elsa.202100179}, pages = {10 Seiten}, year = {2021}, abstract = {The coupling of ligand-stabilized gold nanoparticles with field-effect devices offers new possibilities for label-free biosensing. In this work, we study the immobilization of aminooctanethiol-stabilized gold nanoparticles (AuAOTs) on the silicon dioxide surface of a capacitive field-effect sensor. The terminal amino group of the AuAOT is well suited for the functionalization with biomolecules. The attachment of the positively-charged AuAOTs on a capacitive field-effect sensor was detected by direct electrical readout using capacitance-voltage and constant capacitance measurements. With a higher particle density on the sensor surface, the measured signal change was correspondingly more pronounced. The results demonstrate the ability of capacitive field-effect sensors for the non-destructive quantitative validation of nanoparticle immobilization. In addition, the electrostatic binding of the polyanion polystyrene sulfonate to the AuAOT-modified sensor surface was studied as a model system for the label-free detection of charged macromolecules. Most likely, this approach can be transferred to the label-free detection of other charged molecules such as enzymes or antibodies.}, language = {en} } @article{HaegerWirgesTanzmannetal.2023, author = {Haeger, Gerrit and Wirges, Jessika and Tanzmann, Nicole and Oyen, Sven and Jolmes, Tristan and Jaeger, Karl-Erich and Sch{\"o}rken, Ulrich and Bongaerts, Johannes and Siegert, Petra}, title = {Chaperone assisted recombinant expression of a mycobacterial aminoacylase in Vibrio natriegens and Escherichia coli capable of N-lauroyl-L-amino acid synthesis}, series = {Microbial Cell Factories}, journal = {Microbial Cell Factories}, number = {22}, publisher = {Springer Nature}, issn = {1475-2859}, doi = {10.1186/s12934-023-02079-1}, pages = {Article number: 77 (2023)}, year = {2023}, abstract = {Background Aminoacylases are highly promising enzymes for the green synthesis of acyl-amino acids, potentially replacing the environmentally harmful Schotten-Baumann reaction. Long-chain acyl-amino acids can serve as strong surfactants and emulsifiers, with application in cosmetic industries. Heterologous expression of these enzymes, however, is often hampered, limiting their use in industrial processes. Results We identified a novel mycobacterial aminoacylase gene from Mycolicibacterium smegmatis MKD 8, cloned and expressed it in Escherichia coli and Vibrio natriegens using the T7 overexpression system. The recombinant enzyme was prone to aggregate as inclusion bodies, and while V. natriegens Vmax™ could produce soluble aminoacylase upon induction with isopropyl β-d-1-thiogalactopyranoside (IPTG), E. coli BL21 (DE3) needed autoinduction with lactose to produce soluble recombinant protein. We successfully conducted a chaperone co-expression study in both organisms to further enhance aminoacylase production and found that overexpression of chaperones GroEL/S enhanced aminoacylase activity in the cell-free extract 1.8-fold in V. natriegens and E. coli. Eventually, E. coli ArcticExpress™ (DE3), which co-expresses cold-adapted chaperonins Cpn60/10 from Oleispira antarctica, cultivated at 12 °C, rendered the most suitable expression system for this aminoacylase and exhibited twice the aminoacylase activity in the cell-free extract compared to E. coli BL21 (DE3) with GroEL/S co-expression at 20 °C. The purified aminoacylase was characterized based on hydrolytic activities, being most stable and active at pH 7.0, with a maximum activity at 70 °C, and stability at 40 °C and pH 7.0 for 5 days. The aminoacylase strongly prefers short-chain acyl-amino acids with smaller, hydrophobic amino acid residues. Several long-chain amino acids were fairly accepted in hydrolysis as well, especially N-lauroyl-L-methionine. To initially evaluate the relevance of this aminoacylase for the synthesis of N-acyl-amino acids, we demonstrated that lauroyl-methionine can be synthesized from lauric acid and methionine in an aqueous system. Conclusion Our results suggest that the recombinant enzyme is well suited for synthesis reactions and will thus be further investigated.}, language = {en} } @article{FalkenbergBottBongaertsetal.2022, author = {Falkenberg, Fabian and Bott, Michael and Bongaerts, Johannes and Siegert, Petra}, title = {Phylogenetic survey of the subtilase family and a data-mining-based search for new subtilisins from Bacillaceae}, series = {Frontiers in Microbiology}, volume = {2022}, journal = {Frontiers in Microbiology}, number = {13}, publisher = {Frontiers}, address = {Lausanne}, issn = {1664-302X}, doi = {10.3389/fmicb.2022.1017978}, pages = {Artikel 13:1017978}, year = {2022}, abstract = {The subtilase family (S8), a member of the clan SB of serine proteases are ubiquitous in all kingdoms of life and fulfil different physiological functions. Subtilases are divided in several groups and especially subtilisins are of interest as they are used in various industrial sectors. Therefore, we searched for new subtilisin sequences of the family Bacillaceae using a data mining approach. The obtained 1,400 sequences were phylogenetically classified in the context of the subtilase family. This required an updated comprehensive overview of the different groups within this family. To fill this gap, we conducted a phylogenetic survey of the S8 family with characterised holotypes derived from the MEROPS database. The analysis revealed the presence of eight previously uncharacterised groups and 13 subgroups within the S8 family. The sequences that emerged from the data mining with the set filter parameters were mainly assigned to the subtilisin subgroups of true subtilisins, high-alkaline subtilisins, and phylogenetically intermediate subtilisins and represent an excellent source for new subtilisin candidates.}, language = {en} }