@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} } @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{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{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{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} }