@article{ArreolaOberlaenderMaetzkowetal.2017, author = {Arreola, Julio and Oberl{\"a}nder, Jan and M{\"a}tzkow, M. and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Surface functionalization for spore-based biosensors with organosilanes}, series = {Electrochimica Acta}, volume = {241}, journal = {Electrochimica Acta}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0013-4686}, doi = {10.1016/j.electacta.2017.04.157}, pages = {237 -- 243}, year = {2017}, abstract = {In the present work, surface functionalization of different sensor materials was studied. Organosilanes are well known to serve as coupling agent for biomolecules or cells on inorganic materials. 3-aminopropyltriethoxysilane (APTES) was used to attach microbiological spores time to an interdigitated sensor surface. The functionality and physical properties of APTES were studied on isolated sensor materials, namely silicon dioxide (SiO2) and platinum (Pt) as well as the combined material on sensor level. A predominant immobilization of spores could be demonstrated on SiO2 surfaces. Additionally, the impedance signal of APTES-functionalized biosensor chips has been investigated.}, language = {en} } @article{BronderPoghossianKeusgenetal.2017, author = {Bronder, Thomas and Poghossian, Arshak and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Label-free detection of double-stranded DNA molecules with polyelectrolyte-modified capacitive field-effect sensors}, series = {tm - Technisches Messen}, volume = {84}, journal = {tm - Technisches Messen}, number = {10}, publisher = {De Gruyter}, address = {Oldenbourg}, doi = {10.1515/teme-2017-0015}, pages = {628 -- 634}, year = {2017}, abstract = {In this study, polyelectrolyte-modified field-effect-based electrolyte-insulator-semiconductor (EIS) devices have been used for the label-free electrical detection of double-stranded deoxyribonucleic acid (dsDNA)molecules. The sensor-chip functionalization with a positively charged polyelectrolyte layer provides the possibility of direct adsorptive binding of negatively charged target DNA oligonucleotides onto theSiO2-chip surface.EIS sensors can be utilized as a tool to detect surface-charge changes; the electrostatic adsorption of oligonucleotides onto the polyelectrolyte layer leads to a measureable surface-potential change. Signals of 39mV have been recorded after the incubation with the oligonucleotide solution. Besides the electrochemical experiments, the successful adsorption of dsDNA onto the polyelectrolyte layer has been verified via fluorescence microscopy. The presented results demonstrate that the signal recording of EISchips, which are modified with a polyelectrolyte layer, canbe used as a favorable approach for a fast, cheap and simple detection method for dsDNA.}, language = {en} } @article{MolinnusPoghossianKeusgenetal.2017, author = {Molinnus, Denise and Poghossian, Arshak and Keusgen, Michael and Katz, Evgeny and Sch{\"o}ning, Michael Josef}, title = {Coupling of Biomolecular Logic Gates with Electronic Transducers: From Single Enzyme Logic Gates to Sense/Act/Treat Chips}, series = {Electroanalysis}, volume = {29}, journal = {Electroanalysis}, number = {8}, publisher = {Wiley}, address = {Weinheim}, issn = {1521-4109}, doi = {10.1002/elan.201700208}, pages = {1840 -- 1849}, year = {2017}, abstract = {The integration of biomolecular logic principles with electronic transducers allows designing novel digital biosensors with direct electrical output, logically triggered drug-release, and closed-loop sense/act/treat systems. This opens new opportunities for advanced personalized medicine in the context of theranostics. In the present work, we will discuss selected examples of recent developments in the field of interfacing enzyme logic gates with electrodes and semiconductor field-effect devices. Special attention is given to an enzyme OR/Reset logic gate based on a capacitive field-effect electrolyte-insulator-semiconductor sensor modified with a multi-enzyme membrane. Further examples are a digital adrenaline biosensor based on an AND logic gate with binary YES/NO output and an integrated closed-loop sense/act/treat system comprising an amperometric glucose sensor, a hydrogel actuator, and an insulin (drug) sensor.}, language = {en} } @article{ArreolaKeusgenSchoening2017, author = {Arreola, Julio and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Effect of O2 plasma on properties of electrolyte-insulator-semiconductor structures}, series = {physica status solidi a : applications and materials sciences}, volume = {214}, journal = {physica status solidi a : applications and materials sciences}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201700025}, pages = {Artikel 1700025}, year = {2017}, abstract = {Prior to immobilization of biomolecules or cells onto biosensor surfaces, the surface must be physically or chemically activated for further functionalization. Organosilanes are a versatile option as they facilitate the immobilization through their terminal groups and also display self-assembly. Incorporating hydroxyl groups is one of the important methods for primary immobilization. This can be done, for example, with oxygen plasma treatment. However, this treatment can affect the performance of the biosensors and this effect is not quite well understood for surface functionalization. In this work, the effect of O2 plasma treatment on EIS sensors was investigated by means of electrochemical characterizations: capacitance-voltage (C-V) and constant capacitance (ConCap) measurements. After O2 plasma treatment, the potential of the EIS sensor dramatically shifts to a more negative value. This was successfully reset by using an annealing process.}, 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{SchoeningBronderWuetal.2017, author = {Sch{\"o}ning, Michael Josef and Bronder, Thomas and Wu, Chunsheng and Scheja, Sabrina and Jessing, Max and Metzger-Boddien, Christoph and Keusgen, Michael and Poghossian, Arshak}, title = {Label-Free DNA Detection with Capacitive Field-Effect Devices—Challenges and Opportunities}, series = {Proceedings}, volume = {1}, journal = {Proceedings}, number = {8}, publisher = {MDPI}, address = {Basel}, issn = {2504-3900}, doi = {10.3390/proceedings1080719}, pages = {Artikel 719}, year = {2017}, abstract = {Field-effect EIS (electrolyte-insulator-semiconductor) sensors modified with a positively charged weak polyelectrolyte layer have been applied for the electrical detection of DNA (deoxyribonucleic acid) immobilization and hybridization by the intrinsic molecular charge. The EIS sensors are able to detect the existence of target DNA amplicons in PCR (polymerase chain reaction) samples and thus, can be used as tool for a quick verification of DNA amplification and the successful PCR process. Due to their miniaturized setup, compatibility with advanced micro- and nanotechnologies, and ability to detect biomolecules by their intrinsic molecular charge, those sensors can serve as possible platform for the development of label-free DNA chips. Possible application fields as well as challenges and limitations will be discussed.}, language = {en} } @article{MolinnusHardtSiegertetal.2018, author = {Molinnus, Denise and Hardt, Gabriel and Siegert, Petra and Willenberg, Holger S. and Poghossian, Arshak and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Detection of Adrenaline in Blood Plasma as Biomarker for Adrenal Venous Sampling}, series = {Electroanalysis}, volume = {30}, journal = {Electroanalysis}, number = {5}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1521-4109}, doi = {10.1002/elan.201800026}, pages = {937 -- 942}, year = {2018}, abstract = {An amperometric bi-enzyme biosensor based on substrate recycling principle for the amplification of the sensor signal has been developed for the detection of adrenaline in blood. Adrenaline can be used as biomarker verifying successful adrenal venous sampling procedure. The adrenaline biosensor has been realized via modification of a galvanic oxygen sensor with a bi-enzyme membrane combining a genetically modified laccase and a pyrroloquinoline quinone-dependent glucose dehydrogenase. The measurement conditions such as pH value and temperature were optimized to enhance the sensor performance. A high sensitivity and a low detection limit of about 0.5-1 nM adrenaline have been achieved in phosphate buffer at pH 7.4, relevant for measurements in blood samples. The sensitivity of the biosensor to other catecholamines such as noradrenaline, dopamine and dobutamine has been studied. Finally, the sensor has been successfully applied for the detection of adrenaline in human blood plasma.}, language = {en} } @article{OberlaenderMayerGreeffetal.2018, author = {Oberl{\"a}nder, Jan and Mayer, Marlena and Greeff, Anton and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Spore-based biosensor to monitor the microbicidal efficacy of gaseous hydrogen peroxide sterilization processes}, series = {Biosensors and Bioelectronics}, volume = {104}, journal = {Biosensors and Bioelectronics}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0956-5663}, doi = {10.1016/j.bios.2017.12.045}, pages = {87 -- 94}, year = {2018}, abstract = {In this work, a spore-based biosensor is evaluated to monitor the microbicidal efficacy of sterilization processes applying gaseous hydrogen peroxide (H2O2). The sensor is based on interdigitated electrode structures (IDEs) that have been fabricated by means of thin-film technologies. Impedimetric measurements are applied to study the effect of sterilization process on spores of Bacillus atrophaeus. This resilient microorganism is commonly used in industry to proof the sterilization efficiency. The sensor measurements are accompanied by conventional microbiological challenge tests, as well as morphological characterizations with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The sensor measurements are correlated with the microbiological test routines. In both methods, namely the sensor-based and microbiological one, a tailing effect has been observed. The results are evaluated and discussed in a three-dimensional calibration plot demonstrating the sensor's suitability to enable a rapid process decision in terms of a successfully performed sterilization.}, language = {en} } @article{MolinnusMuschallikGonzalezetal.2018, author = {Molinnus, Denise and Muschallik, Lukas and Gonzalez, Laura Osorio and Bongaerts, Johannes and Wagner, Torsten and Selmer, Thorsten and Siegert, Petra and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Development and characterization of a field-effect biosensor for the detection of acetoin}, series = {Biosensors and Bioelectronics}, volume = {115}, journal = {Biosensors and Bioelectronics}, publisher = {Elsevier}, address = {Amsterdam}, doi = {10.1016/j.bios.2018.05.023}, pages = {1 -- 6}, year = {2018}, abstract = {A capacitive electrolyte-insulator-semiconductor (EIS) field-effect biosensor for acetoin detection has been presented for the first time. The EIS sensor consists of a layer structure of Al/p-Si/SiO₂/Ta₂O₅/enzyme acetoin reductase. The enzyme, also referred to as butane-2,3-diol dehydrogenase from B. clausii DSM 8716T, has been recently characterized. The enzyme catalyzes the (R)-specific reduction of racemic acetoin to (R,R)- and meso-butane-2,3-diol, respectively. Two different enzyme immobilization strategies (cross-linking by using glutaraldehyde and adsorption) have been studied. Typical biosensor parameters such as optimal pH working range, sensitivity, hysteresis, linear concentration range and long-term stability have been examined by means of constant-capacitance (ConCap) mode measurements. Furthermore, preliminary experiments have been successfully carried out for the detection of acetoin in diluted white wine samples.}, language = {en} } @article{MolinnusHardtKaeveretal.2018, author = {Molinnus, Denise and Hardt, G. and K{\"a}ver, L. and Willenberg, H.S. and Kr{\"o}ger, J.-C. and Poghossian, Arshak and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Chip-based biosensor for the detection of low adrenaline concentrations to support adrenal venous sampling}, series = {Sensor and Actuators B: Chemical}, volume = {272}, journal = {Sensor and Actuators B: Chemical}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005}, doi = {10.1016/j.snb.2018.05.136}, pages = {21 -- 27}, year = {2018}, abstract = {A chip-based amperometric biosensor referring on using the bioelectrocatalytical amplification principle for the detection of low adrenaline concentrations is presented. The adrenaline biosensor has been prepared by modification of a platinum thin-film electrode with an enzyme membrane containing the pyrroloquinoline quinone-dependent glucose dehydrogenase and glutaraldehyde. Measuring conditions such as temperature, pH value, and glucose concentration have been optimized to achieve a high sensitivity and a low detection limit of about 1 nM adrenaline measured in phosphate buffer at neutral pH value. The response of the biosensor to different catecholamines has also been proven. Long-term stability of the adrenaline biosensor has been studied over 10 days. In addition, the biosensor has been successfully applied for adrenaline detection in human blood plasma for future biomedical applications. Furthermore, preliminary experiments have been carried to detect the adrenaline-concentration difference measured in peripheral blood and adrenal venous blood, representing the adrenal vein sampling procedure of a physician.}, language = {en} } @article{BronderJessingPoghossianetal.2018, author = {Bronder, Thomas and Jessing, Max P. and Poghossian, Arshak and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Detection of PCR-Amplified Tuberculosis DNA Fragments with Polyelectrolyte-Modified Field-Effect Sensors}, series = {Analytical Chemistry}, volume = {90}, journal = {Analytical Chemistry}, number = {12}, publisher = {ACS Publications}, address = {Washington, DC}, issn = {0003-2700}, doi = {10.1021/acs.analchem.8b01807}, pages = {7747 -- 7753}, year = {2018}, abstract = {Field-effect-based electrolyte-insulator-semiconductor (EIS) sensors were modified with a bilayer of positively charged weak polyelectrolyte (poly(allylamine hydrochloride) (PAH)) and probe single-stranded DNA (ssDNA) and are used for the detection of complementary single-stranded target DNA (cDNA) in different test solutions. The sensing mechanism is based on the detection of the intrinsic molecular charge of target cDNA molecules after the hybridization event between cDNA and immobilized probe ssDNA. The test solutions contain synthetic cDNA oligonucleotides (with a sequence of tuberculosis mycobacteria genome) or PCR-amplified DNA (which origins from a template DNA strand that has been extracted from Mycobacterium avium paratuberculosis-spiked human sputum samples), respectively. Sensor responses up to 41 mV have been measured for the test solutions with DNA, while only small signals of ∼5 mV were detected for solutions without DNA. The lower detection limit of the EIS sensors was ∼0.3 nM, and the sensitivity was ∼7.2 mV/decade. Fluorescence experiments using SybrGreen I fluorescence dye support the electrochemical results.}, language = {en} } @article{JildehOberlaenderKirchneretal.2018, author = {Jildeh, Zaid B. and Oberl{\"a}nder, Jan and Kirchner, Patrick and Keusgen, Michael and Wagner, Patrick H. and Sch{\"o}ning, Michael Josef}, title = {Experimental and Numerical Analyzes of a Sensor Based on Interdigitated Electrodes for Studying Microbiological Alterations}, series = {physica status solidi (a): applications and materials science}, volume = {215}, journal = {physica status solidi (a): applications and materials science}, number = {15}, publisher = {Wiley-VCH}, address = {Weinheim}, issn = {1862-6319}, doi = {10.1002/pssa.201700920}, pages = {Artikel 1700920}, year = {2018}, abstract = {In this work, a cell-based biosensor to evaluate the sterilization efficacy of hydrogen peroxide vapor sterilization processes is characterized. The transducer of the biosensor is based on interdigitated gold electrodes fabricated on an inert glass substrate. Impedance spectroscopy is applied to evaluate the sensor behavior and the alteration of test microorganisms due to the sterilization process. These alterations are related to changes in relative permittivity and electrical conductivity of the bacterial spores. Sensor measurements are conducted with and without bacterial spores (Bacillus atrophaeus), as well as after an industrial sterilization protocol. Equivalent two-dimensional numerical models based on finite element method of the periodic finger structures of the interdigitated gold electrodes are designed and validated using COMSOL® Multiphysics software by the application of known dielectric properties. The validated models are used to compute the electrical properties at different sensor states (blank, loaded with spores, and after sterilization). As a final result, we will derive and tabulate the frequency-dependent electrical parameters of the spore layer using a novel model that combines experimental data with numerical optimization techniques.}, 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{ArreolaKeusgenSchoening2019, author = {Arreola, Julio and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Toward an immobilization method for spore-based biosensors in oxidative environment}, series = {Electrochimica Acta}, volume = {302}, journal = {Electrochimica Acta}, publisher = {Elsevier}, address = {Amsterdam}, doi = {10.1016/j.electacta.2019.01.148}, pages = {394 -- 401}, year = {2019}, language = {en} } @article{BronderPoghossianJessingetal.2019, author = {Bronder, Thomas and Poghossian, Arshak and Jessing, Max P. and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Surface regeneration and reusability of label-free DNA biosensors based on weak polyelectrolyte-modified capacitive field-effect structures}, series = {Biosensors and Bioelectronics}, volume = {126}, journal = {Biosensors and Bioelectronics}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0956-5663}, doi = {10.1016/j.bios.2018.11.019}, pages = {510 -- 517}, year = {2019}, language = {en} } @article{ArreolaKeusgenWagneretal.2019, author = {Arreola, Julio and Keusgen, Michael and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Combined calorimetric gas- and spore-based biosensor array for online monitoring and sterility assurance of gaseous hydrogen peroxide in aseptic filling machines}, series = {Biosensors and Bioelectronics}, volume = {143}, journal = {Biosensors and Bioelectronics}, number = {111628}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0956-5663}, doi = {10.1016/j.bios.2019.111628}, year = {2019}, language = {en} } @article{PilasSelmerKeusgenetal.2019, author = {Pilas, Johanna and Selmer, Thorsten and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Screen-printed carbon electrodes modified with graphene oxide for the design of a reagent-free NAD+-dependent biosensor array}, series = {Analytical Chemistry}, volume = {91}, journal = {Analytical Chemistry}, number = {23}, publisher = {ACS Publications}, address = {Washington}, doi = {10.1021/acs.analchem.9b04481}, pages = {15293 -- 15299}, year = {2019}, 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} }