@article{KarschuckPoghossianSeretal.2024, author = {Karschuck, Tobias and Poghossian, Arshak and Ser, Joey and Tsokolakyan, Astghik and Achtsnicht, Stefan and Wagner, Patrick and Sch{\"o}ning, Michael Josef}, title = {Capacitive model of enzyme-modified field-effect biosensors: Impact of enzyme coverage}, series = {Sensors and Actuators B: Chemical}, volume = {408}, journal = {Sensors and Actuators B: Chemical}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0925-4005 (Print)}, doi = {10.1016/j.snb.2024.135530}, pages = {12 Seiten}, year = {2024}, abstract = {Electrolyte-insulator-semiconductor capacitors (EISCAP) belong to field-effect sensors having an attractive transducer architecture for constructing various biochemical sensors. In this study, a capacitive model of enzyme-modified EISCAPs has been developed and the impact of the surface coverage of immobilized enzymes on its capacitance-voltage and constant-capacitance characteristics was studied theoretically and experimentally. The used multicell arrangement enables a multiplexed electrochemical characterization of up to sixteen EISCAPs. Different enzyme coverages have been achieved by means of parallel electrical connection of bare and enzyme-covered single EISCAPs in diverse combinations. As predicted by the model, with increasing the enzyme coverage, both the shift of capacitance-voltage curves and the amplitude of the constant-capacitance signal increase, resulting in an enhancement of analyte sensitivity of the EISCAP biosensor. In addition, the capability of the multicell arrangement with multi-enzyme covered EISCAPs for sequentially detecting multianalytes (penicillin and urea) utilizing the enzymes penicillinase and urease has been experimentally demonstrated and discussed.}, language = {en} } @article{YoshinobuMiyamotoWagneretal.2024, author = {Yoshinobu, Tatsuo and Miyamoto, Ko-ichiro and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Field-effect sensors combined with the scanned light pulse technique: from artificial olfactory images to chemical imaging technologies}, series = {Chemosensors}, volume = {12}, journal = {Chemosensors}, number = {2}, publisher = {MDPI}, address = {Basel}, issn = {2227-9040}, doi = {10.3390/chemosensors12020020}, pages = {Artikel 20}, year = {2024}, abstract = {The artificial olfactory image was proposed by Lundstr{\"o}m et al. in 1991 as a new strategy for an electronic nose system which generated a two-dimensional mapping to be interpreted as a fingerprint of the detected gas species. The potential distribution generated by the catalytic metals integrated into a semiconductor field-effect structure was read as a photocurrent signal generated by scanning light pulses. The impact of the proposed technology spread beyond gas sensing, inspiring the development of various imaging modalities based on the light addressing of field-effect structures to obtain spatial maps of pH distribution, ions, molecules, and impedance, and these modalities have been applied in both biological and non-biological systems. These light-addressing technologies have been further developed to realize the position control of a faradaic current on the electrode surface for localized electrochemical reactions and amperometric measurements, as well as the actuation of liquids in microfluidic devices.}, language = {en} } @article{BertzSchoeningMolinnusetal.2024, author = {Bertz, Morten and Sch{\"o}ning, Michael Josef and Molinnus, Denise and Homma, Takayuki}, title = {Influence of temperature, light, and H₂O₂ concentration on microbial spore inactivation: in-situ Raman spectroscopy combined with optical trapping}, series = {Physica status solidi (a) applications and materials science}, journal = {Physica status solidi (a) applications and materials science}, number = {Early View}, publisher = {Wiley-VCH}, address = {Berlin}, issn = {1862-6319 (Online)}, doi = {10.1002/pssa.202300866}, pages = {8 Seiten}, year = {2024}, abstract = {To gain insight on chemical sterilization processes, the influence of temperature (up to 70 °C), intense green light, and hydrogen peroxide (H₂O₂) concentration (up to 30\% in aqueous solution) on microbial spore inactivation is evaluated by in-situ Raman spectroscopy with an optical trap. Bacillus atrophaeus is utilized as a model organism. Individual spores are isolated and their chemical makeup is monitored under dynamically changing conditions (temperature, light, and H₂O₂ concentration) to mimic industrially relevant process parameters for sterilization in the field of aseptic food processing. While isolated spores in water are highly stable, even at elevated temperatures of 70 °C, exposure to H₂O₂ leads to a loss of spore integrity characterized by the release of the key spore biomarker dipicolinic acid (DPA) in a concentration-dependent manner, which indicates damage to the inner membrane of the spore. Intensive light or heat, both of which accelerate the decomposition of H₂O₂ into reactive oxygen species (ROS), drastically shorten the spore lifetime, suggesting the formation of ROS as a rate-limiting step during sterilization. It is concluded that Raman spectroscopy can deliver mechanistic insight into the mode of action of H₂O₂-based sterilization and reveal the individual contributions of different sterilization methods acting in tandem.}, language = {en} } @article{EngelmannSimsekShalabyetal.2024, author = {Engelmann, Ulrich M. and Simsek, Beril and Shalaby, Ahmed and Krause, Hans-Joachim}, title = {Key contributors to signal generation in frequency mixing magnetic detection (FMMD): an in silico study}, series = {Sensors}, volume = {24}, journal = {Sensors}, number = {6}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s24061945}, pages = {Artikel 1945}, year = {2024}, abstract = {Frequency mixing magnetic detection (FMMD) is a sensitive and selective technique to detect magnetic nanoparticles (MNPs) serving as probes for binding biological targets. Its principle relies on the nonlinear magnetic relaxation dynamics of a particle ensemble interacting with a dual frequency external magnetic field. In order to increase its sensitivity, lower its limit of detection and overall improve its applicability in biosensing, matching combinations of external field parameters and internal particle properties are being sought to advance FMMD. In this study, we systematically probe the aforementioned interaction with coupled N{\´e}el-Brownian dynamic relaxation simulations to examine how key MNP properties as well as applied field parameters affect the frequency mixing signal generation. It is found that the core size of MNPs dominates their nonlinear magnetic response, with the strongest contributions from the largest particles. The drive field amplitude dominates the shape of the field-dependent response, whereas effective anisotropy and hydrodynamic size of the particles only weakly influence the signal generation in FMMD. For tailoring the MNP properties and parameters of the setup towards optimal FMMD signal generation, our findings suggest choosing large particles of core sizes dc > 25 nm nm with narrow size distributions (σ < 0.1) to minimize the required drive field amplitude. This allows potential improvements of FMMD as a stand-alone application, as well as advances in magnetic particle imaging, hyperthermia and magnetic immunoassays.}, language = {en} } @article{WendlandtKochBritzetal.2023, author = {Wendlandt, Tim and Koch, Claudia and Britz, Beate and Liedek, Anke and Schmidt, Nora and Werner, Stefan and Gleba, Yuri and Vahidpour, Farnoosh and Welden, Melanie and Poghossian, Arshak and Sch{\"o}ning, Michael Josef}, title = {Facile Purification and Use of Tobamoviral Nanocarriers for Antibody-Mediated Display of a Two-Enzyme System}, series = {Viruses}, volume = {9}, journal = {Viruses}, number = {15}, publisher = {MDPI}, address = {Basel}, issn = {1999-4915}, doi = {doi.org/10.3390/v15091951}, pages = {Artikel 1951}, year = {2023}, abstract = {Immunosorbent turnip vein clearing virus (TVCV) particles displaying the IgG-binding domains D and E of Staphylococcus aureus protein A (PA) on every coat protein (CP) subunit (TVCVPA) were purified from plants via optimized and new protocols. The latter used polyethylene glycol (PEG) raw precipitates, from which virions were selectively re-solubilized in reverse PEG concentration gradients. This procedure improved the integrity of both TVCVPA and the wild-type subgroup 3 tobamovirus. TVCVPA could be loaded with more than 500 IgGs per virion, which mediated the immunocapture of fluorescent dyes, GFP, and active enzymes. Bi-enzyme ensembles of cooperating glucose oxidase and horseradish peroxidase were tethered together on the TVCVPA carriers via a single antibody type, with one enzyme conjugated chemically to its Fc region, and the other one bound as a target, yielding synthetic multi-enzyme complexes. In microtiter plates, the TVCVPA-displayed sugar-sensing system possessed a considerably increased reusability upon repeated testing, compared to the IgG-bound enzyme pair in the absence of the virus. A high coverage of the viral adapters was also achieved on Ta2O5 sensor chip surfaces coated with a polyelectrolyte interlayer, as a prerequisite for durable TVCVPA-assisted electrochemical biosensing via modularly IgG-assembled sensor enzymes.}, language = {en} } @article{BertzMolinnusSchoeningetal.2023, author = {Bertz, Morten and Molinnus, Denise and Sch{\"o}ning, Michael Josef and Homma, Takayuki}, title = {Real-time monitoring of H₂O₂ sterilization on individual bacillus atrophaeus spores by optical sensing with trapping Raman spectroscopy}, series = {Chemosensors}, volume = {8}, journal = {Chemosensors}, number = {11}, publisher = {MDPI}, address = {Basel}, issn = {2227-9040}, doi = {10.3390/chemosensors11080445}, pages = {Artikel 445}, year = {2023}, abstract = {Hydrogen peroxide (H₂O₂), a strong oxidizer, is a commonly used sterilization agent employed during aseptic food processing and medical applications. To assess the sterilization efficiency with H₂O₂, bacterial spores are common microbial systems due to their remarkable robustness against a wide variety of decontamination strategies. Despite their widespread use, there is, however, only little information about the detailed time-resolved mechanism underlying the oxidative spore death by H₂O₂. In this work, we investigate chemical and morphological changes of individual Bacillus atrophaeus spores undergoing oxidative damage using optical sensing with trapping Raman microscopy in real-time. The time-resolved experiments reveal that spore death involves two distinct phases: (i) an initial phase dominated by the fast release of dipicolinic acid (DPA), a major spore biomarker, which indicates the rupture of the spore's core; and (ii) the oxidation of the remaining spore material resulting in the subsequent fragmentation of the spores' coat. Simultaneous observation of the spore morphology by optical microscopy corroborates these mechanisms. The dependence of the onset of DPA release and the time constant of spore fragmentation on H₂O₂ shows that the formation of reactive oxygen species from H₂O₂ is the rate-limiting factor of oxidative spore death.}, language = {en} } @article{MoraisSumanSchoeningetal.2023, author = {Morais, Paulo V. and Suman, Pedro H. and Sch{\"o}ning, Michael Josef and Siqueira Junior, Jos{\´e} R. and Orlandi, Marcelo O.}, title = {Layer-by-layer film based on Sn₃O₄ nanobelts as sensing units to detect heavy metals using a capacitive field-effect sensor platform}, series = {Chemosensors}, volume = {11}, journal = {Chemosensors}, number = {8}, publisher = {MDPI}, address = {Basel}, issn = {2227-9040}, doi = {10.3390/chemosensors11080436}, pages = {Artikel 436}, year = {2023}, abstract = {Lead and nickel, as heavy metals, are still used in industrial processes, and are classified as "environmental health hazards" due to their toxicity and polluting potential. The detection of heavy metals can prevent environmental pollution at toxic levels that are critical to human health. In this sense, the electrolyte-insulator-semiconductor (EIS) field-effect sensor is an attractive sensing platform concerning the fabrication of reusable and robust sensors to detect such substances. This study is aimed to fabricate a sensing unit on an EIS device based on Sn₃O₄ nanobelts embedded in a polyelectrolyte matrix of polyvinylpyrrolidone (PVP) and polyacrylic acid (PAA) using the layer-by-layer (LbL) technique. The EIS-Sn₃O₄ sensor exhibited enhanced electrochemical performance for detecting Pb²⁺ and Ni²⁺ ions, revealing a higher affinity for Pb²⁺ ions, with sensitivities of ca. 25.8 mV/decade and 2.4 mV/decade, respectively. Such results indicate that Sn₃O₄ nanobelts can contemplate a feasible proof-of-concept capacitive field-effect sensor for heavy metal detection, envisaging other future studies focusing on environmental monitoring.}, language = {en} } @techreport{HaegerBongaertsSiegert2023, author = {Haeger, Gerrit and Bongaerts, Johannes and Siegert, Petra}, title = {Abschlussbericht Teil II: Eingehende Darstellung Neue biobasierte Lipopeptide aus nachhaltiger Produktion (LipoPep)}, pages = {17Seiten}, year = {2023}, language = {de} } @article{RabehiGarlanAchtsnichtetal.2018, author = {Rabehi, Amine and Garlan, Benjamin and Achtsnicht, Stefan and Krause, Hans-Joachim and Offenh{\"a}usser, Andreas and Ngo, Kieu and Neveu, Sophie and Graff-Dubois, Stephanie and Kokabi, Hamid}, title = {Magnetic detection structure for Lab-on-Chip applications based on the frequency mixing technique}, series = {Sensors}, volume = {18}, journal = {Sensors}, number = {6}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s18061747}, pages = {14 Seiten}, year = {2018}, abstract = {A magnetic frequency mixing technique with a set of miniaturized planar coils was investigated for use with a completely integrated Lab-on-Chip (LoC) pathogen sensing system. The system allows the detection and quantification of superparamagnetic beads. Additionally, in terms of magnetic nanoparticle characterization ability, the system can be used for immunoassays using the beads as markers. Analytical calculations and simulations for both excitation and pick-up coils are presented; the goal was to investigate the miniaturization of simple and cost-effective planar spiral coils. Following these calculations, a Printed Circuit Board (PCB) prototype was designed, manufactured, and tested for limit of detection, linear response, and validation of theoretical concepts. Using the magnetic frequency mixing technique, a limit of detection of 15 µg/mL of 20 nm core-sized nanoparticles was achieved without any shielding.}, language = {en} } @article{AchtsnichtSchoenenbornOffenhaeusseretal.2019, author = {Achtsnicht, Stefan and Sch{\"o}nenborn, Kristina and Offenh{\"a}usser, Andreas and Krause, Hans-Joachim}, title = {Measurement of the magnetophoretic velocity of different superparamagnetic beads}, series = {Journal of Magnetism and Magnetic Materials}, volume = {477}, journal = {Journal of Magnetism and Magnetic Materials}, number = {1}, publisher = {Elsevier}, address = {Amsterdam}, issn = {0304-8853}, doi = {10.1016/j.jmmm.2018.10.066}, pages = {244 -- 248}, year = {2019}, abstract = {The movement of magnetic beads due to a magnetic field gradient is of great interest in different application fields. In this report we present a technique based on a magnetic tweezers setup to measure the velocity factor of magnetically actuated individual superparamagnetic beads in a fluidic environment. Several beads can be tracked simultaneously in order to gain and improve statistics. Furthermore we show our results for different beads with hydrodynamic diameters between 200 and 1000 nm from diverse manufacturers. These measurement data can, for example, be used to determine design parameters for a magnetic separation system, like maximum flow rate and minimum separation time, or to select suitable beads for fixed experimental requirements.}, language = {en} } @article{AchtsnichtToedterNiehuesetal.2019, author = {Achtsnicht, Stefan and T{\"o}dter, Julia and Niehues, Julia and Tel{\"o}ken, Matthias and Offenh{\"a}usser, Andreas and Krause, Hans-Joachim and Schr{\"o}per, Florian}, title = {3D printed modular immunofiltration columns for frequency mixing-based multiplex magnetic immunodetection}, series = {Sensors}, volume = {19}, journal = {Sensors}, number = {1}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s19010148}, pages = {15 Seiten}, year = {2019}, abstract = {For performing point-of-care molecular diagnostics, magnetic immunoassays constitute a promising alternative to established enzyme-linked immunosorbent assays (ELISA) because they are fast, robust and sensitive. Simultaneous detection of multiple biomolecular targets from one body fluid sample is desired. The aim of this work is to show that multiplex magnetic immunodetection based on magnetic frequency mixing by means of modular immunofiltration columns prepared for different targets is feasible. By calculations of the magnetic response signal, the required spacing between the modules was determined. Immunofiltration columns were manufactured by 3D printing and antibody immobilization was performed in a batch approach. It was shown experimentally that two different target molecules in a sample solution could be individually detected in a single assaying step with magnetic measurements of the corresponding immobilization filters. The arrangement order of the filters and of a negative control did not influence the results. Thus, a simple and reliable approach to multi-target magnetic immunodetection was demonstrated.}, language = {en} } @article{AchtsnichtPourshahidiOffenhaeusseretal.2019, author = {Achtsnicht, Stefan and Pourshahidi, Ali Mohammad and Offenh{\"a}usser, Andreas and Krause, Hans-Joachim}, title = {Multiplex detection of different magnetic beads using frequency scanning in magnetic frequency mixing technique}, series = {Sensors}, volume = {19}, journal = {Sensors}, number = {11}, publisher = {MDPI}, address = {Basel}, issn = {1424-8220}, doi = {10.3390/s19112599}, pages = {13 Seiten}, year = {2019}, abstract = {In modern bioanalytical methods, it is often desired to detect several targets in one sample within one measurement. Immunological methods including those that use superparamagnetic beads are an important group of techniques for these applications. The goal of this work is to investigate the feasibility of simultaneously detecting different superparamagnetic beads acting as markers using the magnetic frequency mixing technique. The frequency of the magnetic excitation field is scanned while the lower driving frequency is kept constant. Due to the particles' nonlinear magnetization, mixing frequencies are generated. To record their amplitude and phase information, a direct digitization of the pickup-coil's signal with subsequent Fast Fourier Transformation is performed. By synchronizing both magnetic beads using frequency scanning in magnetic frequency mixing technique magnetic fields, a stable phase information is gained. In this research, it is shown that the amplitude of the dominant mixing component is proportional to the amount of superparamagnetic beads inside a sample. Additionally, it is shown that the phase does not show this behaviour. Excitation frequency scans of different bead types were performed, showing different phases, without correlation to their diverse amplitudes. Two commercially available beads were selected and a determination of their amount in a mixture is performed as a demonstration for multiplex measurements.}, language = {en} } @article{AchtsnichtNeuendorfFassbenderetal.2019, author = {Achtsnicht, Stefan and Neuendorf, Christian and Faßbender, Tobias and N{\"o}lke, Greta and Offenh{\"a}usser, Andreas and Krause, Hans-Joachim and Schr{\"o}per, Florian}, title = {Sensitive and rapid detection of cholera toxin subunit B using magnetic frequency mixing detection}, series = {Plos One}, volume = {14}, journal = {Plos One}, number = {7}, publisher = {Plos}, address = {San Francisco}, issn = {1932-6203}, doi = {10.1371/journal.pone.0219356}, pages = {e0219356}, year = {2019}, abstract = {Cholera is a life-threatening disease caused by the cholera toxin (CT) as produced by some Vibrio cholerae serogroups. In this research we present a method which directly detects the toxin's B subunit (CTB) in drinking water. For this purpose we performed a magnetic sandwich immunoassay inside a 3D immunofiltration column. We used two different commercially available antibodies to capture CTB and for binding to superparamagnetic beads. ELISA experiments were performed to select the antibody combination. The beads act as labels for the magnetic frequency mixing detection technique. We show that the limit of detection depends on the type of magnetic beads. A nonlinear Hill curve was fitted to the calibration measurements by means of a custom-written python software. We achieved a sensitive and rapid detection of CTB within a broad concentration range from 0.2 ng/ml to more than 700 ng/ml.}, language = {en} } @phdthesis{Achtsnicht2020, author = {Achtsnicht, Stefan}, title = {Multiplex-Magnetdetektion von superparamagnetischen Beads zur Identifizierung von Trinkwasserkontaminationen}, doi = {10.18154/RWTH-2020-12052}, pages = {144 Seiten}, year = {2020}, abstract = {Die qualitative und quantitative Detektion von Zielsubstanzen innerhalb einer w{\"a}ssrigen Probe ist f{\"u}r viele Fragestellungen von Interesse, etwa bei der Detektion von Kontaminationen in Trinkwasser in Krisensituationen. Hierbei ist es nicht nur wichtig, dass Pathogene m{\"o}glichst sensitiv detektiert werden k{\"o}nnen, sondern auch, dass die Analyse schnell erfolgt, um Betroffenen im Katastrophenfall z{\"u}gig sicheres Trinkwasser zu Verf{\"u}gung stellen zu k{\"o}nnen. Da bei einem solchen Szenario nicht von einer in der N{\"a}he befindlichen funktionierenden Laborinfrastruktur ausgegangen werden kann, ist es wichtig, dass die Messung direkt vor Ort erfolgen kann. Im Rahmen dieser Arbeit wurde untersucht, ob eine derartige Schnellanalytik mithilfe von superparamagnetischen Beads (MBs) und der magnetischen Frequenzmischtechnik m{\"o}glich ist. Dabei werden die MBs mit Hilfe von prim{\"a}ren Antik{\"o}rpern an die Zielsubstanz gebunden und mit sekund{\"a}ren Antik{\"o}rpern an die Poren-Oberfl{\"a}che eines Polyethylen-Filters fixiert (Sandwich-Immunoassay). So kann die Quantifizierung der Zielsubstanz auf eine magnetische Messung der immobilisierten MB-Marker zur{\"u}ckgef{\"u}hrt werden. Die magnetische Frequenzmischtechnik basiert auf der Anregung der Probe mit Magnetfeldern zweier verschiedener Frequenzen. Die durch die nichtlineare Magnetisierungsform der superparamagnetischen MBs entstehenden Mischfrequenzen werden typischerweise mithilfe einer zweistufigen Lock-in-Detektion analysiert (analoge Demodulation), die in einem Magnetreader als Handheldger{\"a}t realisiert wurde. Zus{\"a}tzlich zu dieser Technik wurde das Prinzip der direkten Digitalisierung des gesamten Antwortsignals mit anschließender Fourier-Analyse der erzeugten Mischfrequenzen experimentell umgesetzt, um die Amplituden und Phasen mehrerer Mischfrequenzen simultan zu erfassen. Eine M{\"o}glichkeit zur Sensitivit{\"a}tssteigerung ist die magnetische Aufkonzentration, indem vor der magnetischen Analyse eine Separation der MBs aus einem gr{\"o}ßeren Probenvolumen mittels magnetischem Feldgradienten durchgef{\"u}hrt wird. Zur Charakterisierung verschiedener kommerzieller MBs hinsichtlich ihrer magnetischen Separierbarkeit wurde ein Aufbau zur Messung ihrer magnetophoretischen Beweglichkeiten realisiert und ihre Geschwindigkeiten im Gradientenfeld mikroskopisch gemessen.Da eine Probe oftmals nicht nur auf eine einzige Zielsubstanz, sondern simultan auf mehrere verschiedene Pathogene hin untersucht werden soll, wurden verschiedene Ans{\"a}tze entwickelt und getestet, die einen solchen multiparametrischen magnetischen Immunoassay erm{\"o}glichen. Einerseits wurde eine r{\"a}umliche Separation der Bindungsbereiche f{\"u}r verschiedene Zielsubstanzen realisiert, die sequentiell ausgewertet werden k{\"o}nnen. Andererseits wurde die Unterscheidung von verschiedenen Zielsubstanzen anhand der Charakteristika der an sie gebundenen, verschieden funktionalisierten MB-Typen untersucht. F{\"u}r eine solche Unterscheidung wurde zum einen die Anregefrequenz der magnetischen Frequenzmischtechnik w{\"a}hrend einer Messung variiert. Damit konnte gezeigt werden, dass sich verschiedene MB-Sorten anhand der Phase ihrer Frequenzmischsignale voneinander unterscheiden lassen. Weiterhin wurde gezeigt, dass sich der Signalverlauf einer bin{\"a}ren Mischung zweier verschiedener MB-Typen als gradueller {\"U}bergang der Verl{\"a}ufe der beiden reinen MB-L{\"o}sungen ergibt. Eine weitere Analysemethode f{\"u}r einen multiparametrischen Immunoassay besteht darin, ein zus{\"a}tzliches einstellbares statisches magnetisches Offsetfeld zu verwenden. Hierf{\"u}r wurden mehrere Aufbauten auf Basis von Permanent- und Elektromagneten simuliert, konstruiert und charakterisiert. Mithilfe von Simulationen konnte gezeigt werden, dass eine auf diesem Verfahren beruhende Unterscheidung f{\"u}r MBs mit unterschiedlichen magnetischen Partikelmomenten m{\"o}glich ist. Als direkte Anwendung des hier entwickelten Magnetreaders in Zusammenspiel mit der digitalen Demodulation wurde ein magnetischer Assay gegen die B-Untereinheit des Choleratoxins in Trinkwasser mit einem niedrigen Detektionslimit von 0,2 ng/ml demonstriert.}, language = {de} } @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{WeldenJablonskiWegeetal.2021, author = {Welden, Rene and Jablonski, Melanie and Wege, Christina and Keusgen, Michael and Wagner, Patrick Hermann and Wagner, Torsten and Sch{\"o}ning, Michael Josef}, title = {Light-Addressable Actuator-Sensor Platform for Monitoring and Manipulation of pH Gradients in Microfluidics: A Case Study with the Enzyme Penicillinase}, series = {Biosensors}, volume = {11}, journal = {Biosensors}, number = {6}, publisher = {MDPI}, address = {Basel}, issn = {2079-6374}, doi = {10.3390/bios11060171}, pages = {Artikel 171}, year = {2021}, abstract = {The feasibility of light-addressed detection and manipulation of pH gradients inside an electrochemical microfluidic cell was studied. Local pH changes, induced by a light-addressable electrode (LAE), were detected using a light-addressable potentiometric sensor (LAPS) with different measurement modes representing an actuator-sensor system. Biosensor functionality was examined depending on locally induced pH gradients with the help of the model enzyme penicillinase, which had been immobilized in the microfluidic channel. The surface morphology of the LAE and enzyme-functionalized LAPS was studied by scanning electron microscopy. Furthermore, the penicillin sensitivity of the LAPS inside the microfluidic channel was determined with regard to the analyte's pH influence on the enzymatic reaction rate. In a final experiment, the LAE-controlled pH inhibition of the enzyme activity was monitored by the LAPS.}, language = {en} } @article{WeldenPoghossianVahidpouretal.2022, author = {Welden, Melanie and Poghossian, Arshak and Vahidpour, Farnoosh and Wendlandt, Tim and Keusgen, Michael and Wege, Christina and Sch{\"o}ning, Michael Josef}, title = {Towards multi-analyte detection with field-effect capacitors modified with tobacco mosaic virus bioparticles as enzyme nanocarriers}, series = {Biosensors}, volume = {12}, journal = {Biosensors}, number = {1}, publisher = {MDPI}, address = {Basel}, issn = {2079-6374}, doi = {10.3390/bios12010043}, pages = {Artikel 43}, year = {2022}, abstract = {Utilizing an appropriate enzyme immobilization strategy is crucial for designing enzyme-based biosensors. Plant virus-like particles represent ideal nanoscaffolds for an extremely dense and precise immobilization of enzymes, due to their regular shape, high surface-to-volume ratio and high density of surface binding sites. In the present work, tobacco mosaic virus (TMV) particles were applied for the co-immobilization of penicillinase and urease onto the gate surface of a field-effect electrolyte-insulator-semiconductor capacitor (EISCAP) with a p-Si-SiO₂-Ta₂O₅ layer structure for the sequential detection of penicillin and urea. The TMV-assisted bi-enzyme EISCAP biosensor exhibited a high urea and penicillin sensitivity of 54 and 85 mV/dec, respectively, in the concentration range of 0.1-3 mM. For comparison, the characteristics of single-enzyme EISCAP biosensors modified with TMV particles immobilized with either penicillinase or urease were also investigated. The surface morphology of the TMV-modified Ta₂O₅-gate was analyzed by scanning electron microscopy. Additionally, the bi-enzyme EISCAP was applied to mimic an XOR (Exclusive OR) enzyme logic gate.}, language = {en} } @inproceedings{WeldenSeverinsPoghossianetal.2022, author = {Welden, Melanie and Severins, Robin and Poghossian, Arshak and Wege, Christina and Siegert, Petra and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Studying the immobilization of acetoin reductase with Tobacco mosaic virus particles on capacitive field-effect sensors}, series = {2022 IEEE International Symposium on Olfaction and Electronic Nose (ISOEN)}, booktitle = {2022 IEEE International Symposium on Olfaction and Electronic Nose (ISOEN)}, publisher = {IEEE}, isbn = {978-1-6654-5860-3 (Online)}, doi = {10.1109/ISOEN54820.2022.9789657}, pages = {4 Seiten}, year = {2022}, abstract = {A capacitive electrolyte-insulator-semiconductor (EISCAP) biosensor modified with Tobacco mosaic virus (TMV) particles for the detection of acetoin is presented. The enzyme acetoin reductase (AR) was immobilized on the surface of the EISCAP using TMV particles as nanoscaffolds. The study focused on the optimization of the TMV-assisted AR immobilization on the Ta 2 O 5 -gate EISCAP surface. The TMV-assisted acetoin EISCAPs were electrochemically characterized by means of leakage-current, capacitance-voltage, and constant-capacitance measurements. The TMV-modified transducer surface was studied via scanning electron microscopy.}, language = {en} } @article{WeldenSeverinsPoghossianetal.2022, author = {Welden, Melanie and Severins, Robin and Poghossian, Arshak and Wege, Christina and Bongaerts, Johannes and Siegert, Petra and Keusgen, Michael and Sch{\"o}ning, Michael Josef}, title = {Detection of acetoin and diacetyl by a tobacco mosaic virus-assisted field-effect biosensor}, series = {Chemosensors}, volume = {10}, journal = {Chemosensors}, number = {6}, publisher = {MDPI}, address = {Basel}, issn = {2227-9040}, doi = {10.3390/chemosensors10060218}, pages = {Artikel 218}, year = {2022}, abstract = {Acetoin and diacetyl have a major impact on the flavor of alcoholic beverages such as wine or beer. Therefore, their measurement is important during the fermentation process. Until now, gas chromatographic techniques have typically been applied; however, these require expensive laboratory equipment and trained staff, and do not allow for online monitoring. In this work, a capacitive electrolyte-insulator-semiconductor sensor modified with tobacco mosaic virus (TMV) particles as enzyme nanocarriers for the detection of acetoin and diacetyl is presented. The enzyme acetoin reductase from Alkalihalobacillus clausii DSM 8716ᵀ is immobilized via biotin-streptavidin affinity, binding to the surface of the TMV particles. The TMV-assisted biosensor is electrochemically characterized by means of leakage-current, capacitance-voltage, and constant capacitance measurements. In this paper, the novel biosensor is studied regarding its sensitivity and long-term stability in buffer solution. Moreover, the TMV-assisted capacitive field-effect sensor is applied for the detection of diacetyl for the first time. The measurement of acetoin and diacetyl with the same sensor setup is demonstrated. Finally, the successive detection of acetoin and diacetyl in buffer and in diluted beer is studied by tuning the sensitivity of the biosensor using the pH value of the measurement solution.}, language = {en} } @article{HaegerBongaertsSiegert2022, author = {Haeger, Gerrit and Bongaerts, Johannes and Siegert, Petra}, title = {A convenient ninhydrin assay in 96-well format for amino acid-releasing enzymes using an air-stable reagent}, series = {Analytical Biochemistry}, journal = {Analytical Biochemistry}, number = {624}, publisher = {Elsevier}, address = {Amsterdam}, issn = {1096-0309}, doi = {10.1016/j.ab.2022.114819}, pages = {Artikel 114819}, year = {2022}, abstract = {An improved and convenient ninhydrin assay for aminoacylase activity measurements was developed using the commercial EZ Nin™ reagent. Alternative reagents from literature were also evaluated and compared. The addition of DMSO to the reagent enhanced the solubility of Ruhemann's purple (RP). Furthermore, we found that the use of a basic, aqueous buffer enhances stability of RP. An acidic protocol for the quantification of lysine was developed by addition of glacial acetic acid. The assay allows for parallel processing in a 96-well format with measurements microtiter plates.}, language = {en} }