TY - CONF A1 - Ecke, Alexander T1 - An Immunosensor for Pharmaceutical Contaminants in Water Supply N2 - Contamination of drinking water with pharmaceuticals is an emerging problem and a potential health threat. Immunochemical methods based on the binding of the analyte to specific antibodies enable fast & cost-effective on-site analyses. Magnetic bead-based immunoassays (MBBAs) allow for implementation into an immunosensor for online testing. Particles are prepared by either coupling the analyte molecule (diclofenac) or the respective antibody (amoxicillin) to the surface. For miniaturization, detection is performed electrochemically (chronoamperometry) on a microfluidic chip. The developed immunosensor will enable detection of pharmaceuticals directly in water supply pipes. T2 - 11th SETAC Young Environmental Scientists Meeting CY - Online meeting DA - 07.03.2022 KW - Immunosensor KW - Magnetic Beads KW - Water Analysis KW - Electrochemistry KW - Microfluidics PY - 2022 AN - OPUS4-54432 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Ecke, Alexander T1 - An Immunosensor for Pharmaceutical Contaminants in Drinking Water N2 - Contamination of drinking water with pharmaceuticals is an emerging problem and a potential health threat. Immunochemical methods based on the binding of the analyte to specific antibodies enable fast & cost-effective on-site analyses. Magnetic bead-based immunoassays (MBBAs) allow for implementation into an immunosensor for online testing. Particles are prepared by either coupling the analyte molecule (diclofenac) or the respective antibody (amoxicillin) to the surface. For miniaturization, detection is performed electrochemically (chronoamperometry) on a microfluidic chip. The developed immunosensor will enable detection of pharmaceuticals directly in water supply pipes. T2 - JCF Frühjahrssymposium 2022 CY - Hannover, Germany DA - 23.03.2022 KW - Immunosensor KW - Electrochemistry KW - Amoxicillin KW - Diclofenac KW - Magnetic Beads PY - 2022 AN - OPUS4-54627 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Rudolf A1 - He, Y. A1 - Zhu, R. A1 - Cai, Y. A1 - Zhang, Y. A1 - Zhang, Y. A1 - Pan, S. A1 - Zhang, Y. T1 - Transcriptomics and protein biomarkers reveal the detoxifying mechanisms of UV radiation for nebivolol toward zebrafish (Danio rerio) embryos/larvae N2 - Nebivolol (NEB), a β-blocker frequently used to treat cardiovascular diseases, has been widely detected in aquatic environments, and can be degraded under exposure to UV radiation, leading to the formation of certain transformation products (UV-TPs). Thus, the toxic effects of NEB and its UV-TPs on aquatic organisms are of great importance for aquatic ecosystems. In the present study, the degradation pathway of NEB under UV radiation was investigated. Subsequently, zebrafish embryos/larvae were used to assess the median lethal concentration (LC50) of NEB, and to clarify the sub-lethal effects of NEB and its UV-TPs for the first time. It was found that UV radiation could reduce the toxic effects of NEB on the early development of zebrafish. Transcriptomic analysis identified the top 20 enriched Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways in zebrafish larvae exposed to NEB, most of which were associated with the antioxidant, nervous, and immune systems. The number of differentially expressed genes (DEGs) in the pathways were reduced after UV radiation. Furthermore, the analysis of protein biomarkers, including CAT and GST (antioxidant response), AChE and ACh (neurotoxicity), CRP and LYS (immune response), revealed that NEB exposure reduced the activity of these biomarkers, whereas UV radiation could alleviate the effects. The present study provides initial insights into the mechanisms underlying toxic effects of NEB and the detoxification effects of UV radiation on the early development of zebrafish. It highlights the necessity of considering the toxicity of UV-TPs when evaluating the toxicity of emerging pollutants in aquatic systems. KW - Biomarker KW - Pharmazeutika KW - Toxikologie KW - UV Bestrahlung KW - Zebrafisch KW - Transformationsprodukte PY - 2022 DO - https://doi.org/10.1016/j.aquatox.2022.106241 SN - 0166-445X VL - 249 SP - 1 EP - 10 PB - Elsevier CY - Amsterdam AN - OPUS4-55559 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Rudolf A1 - Almeida, Â. A1 - Calisto, V. A1 - Esteves, V. I. A1 - Soares, A. M. V. M. A1 - Freitas, R. T1 - Salinity-dependent impacts on the effects of antiepileptic and antihistaminic drugs in Ruditapes philippinarum N2 - In Coastal Systems, pollutants as pharmaceutical drugs exert changes from the molecular to the organism level in marine bivalves. Besides pollutants, Coastal Systems are prone to changes in environmental Parameters, as the alteration of salinity values because of Climate Change. Together, these Stressors (pharmaceutical drugs and salinity changes) can exert different threats than each Stressor acting individually; for example, salinity can change the physical-chemical properties of the drugs and/or the sensitivity of the organisms to them. However, limited Information is available on this subject, with variable results, and for this reason, this study aimed to evaluate the impacts of salinity changes (15,25 and 35) on the effects of the antiepileptic carbamazepine (CBZ, 1 (ig/L) and the antihistamine cetirizine (CTZ, 0.6 pg/L), when acting individually and combined (CBZ + CTZ), in the edible clam Ruditapes philippinarum. After 28 days ofexposure, drugs concentrations, bioconcentration factors and biochemical parameters, related to clam's metabolic caparity and oxidative stress were evaluated. The results showed that dams under low salinity suffered more changes in metabolic, antioxidant and biotransformation activities, in comparison with the remaining salinities under study. However, limited impacts were observed when comparing drug effects at low salinity. Indeed, it seemed that CTZ and CBZ + CTZ, under high salinity (salinity 35) were the worst exposure conditions for the dams, since they caused higher leveis of cellular damage. It Stands out that salinity changes altered the impact of pharmaceutical drugs on marine bivalves. KW - Muscheln KW - Salinität KW - Carbamazepin KW - Cetirizin KW - ELISA KW - Immunoassay KW - Antiepileptikum PY - 2022 DO - https://doi.org/10.1016/j.scitotenv.2021.150369 SN - 1879-1026 VL - 806 SP - 1 EP - 13 PB - Elsevier Science CY - Amsterdam AN - OPUS4-55561 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Almeida, Â. A1 - Calisto, V. A1 - Esteves, V. I. A1 - Schneider, Rudolf A1 - Soares, A. M. V. M. A1 - Freitas, R. T1 - Responses of Ruditapes philippinarum to contamination by pharmaceutical drugs under ocean acidification scenario N2 - In coastal systems, organisms are exposed to amultitude of stressors whose interactions and effects are poorly studied. Pharmaceutical drugs and Climate Change consequences, such as lowered pH, are examples of stressors affecting marine organisms, as bivalves. Although a vast literature is available for the effects of these stressors when acting individually, very limited information exists on the impacts that the combination of both can have on marine bivalves. For this reason, this study aimed to evaluate the impacts of a simulated ocean acidification scenario (control pH, 8.0; lowered pH, pH 7.6) on the effects of the antiepileptic carbamazepine (CBZ, 1 μg/L) and the antihistamine cetirizine (CTZ, 0.6 μg/L), when acting individually and combined (CBZ + CTZ), on the edible clam Ruditapes philippinarum. After 28 days of exposure, drug concentrations, bioconcentration factors and biochemical parameters related to the clams' metabolic capacity and oxidative stress were evaluated. The results showed that R. philippinarum clams responded differently to pharmaceutical drugs depending on the pH tested, influencing both bioconcentration and biological responses. In general, drug combined treatments showed fewer impacts than drugs acting alone, and acidification seemed to activate at a higher extension the elimination processes that were not activated under control pH. Also, lowered pH per se exerted negative impacts (e.g., cellular damage) on R. philippinarum and the combination with pharmaceutical drugs did not enhance the toxicity. KW - Biosensoren KW - Immunoassay KW - ELISA KW - Vor-Ort-Analytik KW - Toxikologie KW - Pharmaceutical drugs KW - Bivalves KW - Ocean acidification KW - Biomarkers KW - Climate change PY - 2022 DO - https://doi.org/10.1016/j.scitotenv.2022.153591 SN - 1879-1026 VL - 824 SP - 1 EP - 11 PB - Elsevier Science CY - Amsterdam AN - OPUS4-55590 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - GEN A1 - Bayram, Rabia T1 - Rekombinante Herstellung einer Amin-Oxidase aus Aspergillus niger und deren Einsatz in der Biosensorik N2 - Das Ziel der vorliegenden Arbeit war die rekombinante Herstellung der AnFAO in E. coli, welche ursprünglich von A. niger produziert wird, und deren anschließende Anwendung in der Entwick-lung eines Fumonisin- Biosensors für die Lebensmittelanalytik. In Bezug auf die aktuell steigen-den Lebensmittel- und Rohstoffpreise ist die Entwicklung eines Schnelltestsystems zur Detektion von Kontaminationen mit Mykotoxinen besonders gefragt, da zusätzliche Verluste und Kosten verhindert werden können. Für die Herstellung der AnFAO als MBP-Fusionsprotein und Biotin-Fusionsprotein mit zusätzlichem 6x His-Tag wurden die pET-MBP-AnFAO und pRSET-BH6-An-FAO Expressionsvektoren erfolgreich konstruiert. Die lösliche Expression im Cytoplasma wurde von MBP-AnFAO erreicht, wobei die Expression von BH6-AnFAO in inclusion bodies erfolgte und keine Aktivität des Enzyms nachgewiesen wurde. Die Enzymaktivität von MBP-AnFAO wurde im optischen Assay und amperometrisch durch die Detektion von H2O2 ermittelt. Im optischen Assay wurde die maximale spezifische Aktivität von 2,45 U/mg bei 45 °C, einer Enzymkonzentration von 3 μM und 50 μM FB1 erreicht. Generell wurden mit FB1 als Enzymsubstrat höhere Fluoreszenzsig-nale und dementsprechend höhere Enzymaktivitäten als für FB2 beobachtet. Es wurde ein linearer Zusammenhang der Enzymaktivitäten mit der Substratkonzentration beobachtet. Außerdem wurde ein Anstieg der H2O2-Konzentration mit zunehmender Inkubationszeit (bis zu 60 Minuten) erkannt. Des Weiteren wurde der Einfluss des MBP-Fusionsproteins auf die Enzymaktivität des AnFAO untersucht. Dabei wurde anhand von wesentlich geringeren Aktivitäten für AnFAO allein ein eventueller Einfluss des MBP auf dessen Aktivität festgestellt. Ein elektrochemischer Aufbau wurde zur Entwicklung eines Fumonisin- Sensors genutzt. Dazu wurde nachgewiesen, dass FB1 von rekombinant hergestelltem MBP-AnFAO unter Bildung von NH3 und H2O2 zu weniger toxi-schem FB1, dem FPy1, oxidiert werden kann. An der PB/Kohlenstoff-Elektrode, welche sensitiver gegenüber H2O2 und stabiler als die CoPC/Kohlenstoff Elektrode war, wurde die Reduktion des enzymatisch gebildeten H2O2 gemessen. Die Erstellung einer FB1- Kalibriergerade mit FB1- Lösun-gen verschiedener Konzentrationen zeigte, dass das Enzym auch in immobilisierter Form in der Lage ist unterschiedliche Konzentrationen an FB1 zu desaminieren. Dazu gehört auch der Kon-zentrationsbereich unterhalb des festgelegten Grenzwertes von 2,7 μmol/kg für Fumonisin. Die amperometrischen Kurven hierfür wurden an der PB/Kohlenstoff-Elektrode erfolgreich aufge-nommen. Störungen des H2O2- Redoxstroms durch das Enzym, der Enzymsubstrate und der Ne-benprodukte während der amperometrischen Messung wurden ausgeschlossen. Sowohl im opti-schen als auch im amperometrischen Assay wurde ausschließlich das enzymatisch gebildete H2O2 bestimmt. KW - Fumonisin KW - Mykotoxin KW - rekombinante Protein Expression KW - Amperometrie PY - 2022 SP - 1 EP - 104 PB - Berliner Hochschule für Technik CY - Berlin AN - OPUS4-57724 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - GEN A1 - Ferreira Camoes Liestmann, Zoe T1 - Development of an electrochemiluminescence immunoassay for selected pathogens in wastewater N2 - The outbreak of SARS-CoV-2 in December of 2019, led to a worldwide still on-going pandemic. Since then, several so-called waves of SARS-CoV-2 infections, a time period with a high and fast rising number of new infections, have occurred all over the world. Classic surveillance approaches are hardly applicable, and further, non-detected cases cannot be covered by them. Wastewater-based Epidemiology (WBE) was proven to be a reliable tool for the prediction of new SARS-CoV-2 infection waves, due to the discharge of virus particles in fecal shedding of infectious people. Until now, for the monitoring of SARS-CoV-2 in wastewater, Polymerase Chain Reaction (PCR) is used as analytical tool. Even though PCR is a highly sensitive analytical tool, is presents several disadvantages, such as the need for trained personnel, specific technical equipment, as well as a difficult performance. An analytical tool, to which these disadvantaged do not apply, are immunoassays. In this work, a sandwich Enzyme-Linked Immunosorbent Assay (ELISA), with the immobilization of the capture antibodies on the surface of a Microtiter Plate (MTP), as well as a sandwich Magnetic Bead-Based Assay (MBBA), with immobilization of the capture antibodies on the surface of Magnetic Beads (MBs), targeting the SARS-CoV-2 N-protein, were developed and optimized. Both assay formats were performed with a colorimetric and chemiluminescent detection. The developed assay is composed of the two monoclonal antibodies (mAb) AH2 and DE6 - which was biotinylated in the course of the work - which bind to two different epitops of the antigen N-protein. As tracer, Neutravidin-HRP was used, which binds, through interaction of the Neutravidin with the biotin, to the mAb DE6-Biotin. The assay development and optimization procedure included the investigation of the surface saturation with the mAb AH2, the concentration and dilution of the mAb DE6-Biotin and Neutravidin-HRP, the ideal MBs, the ideal coating as well as dilution buffers, and the colorimetric and chemiluminescent substrates. For the developed and fully optimized colorimetric ELISA, a test midpoint x0 of 388 μg/L, for the chemiluminsecent ELISA of 371 μg/L, for the colorimetric MBBA of 251 μg/L and for the chemiluminescent MBBA of 243 μg/L was obtained. Validation of the colorimetric MBBA was done by measurement of three wastewater samples collected at the Wastewater Treatment Plant (WWTP) Potsdam. Whilst no N-protein could be detected in the samples, by spiking of the wastewater samples with certain concentrations of the N-protein, 10- to 18-times lower concentrations could be back-calculated, which can be attributed to matrix-effects of the wastewater sample. Next to the matrix-effects, also several other reason exist, why no N-protein could be determined in the samples. Because of that, further investigation of the handling, and the measurement of the wastewater samples, as well as the improvement of the assay sensitivity through further optimization steps or exchange of the antibodies, is still necessary. KW - SARS-CoV-2 KW - ELISA KW - Antibody KW - N-capsid PY - 2022 SP - 1 EP - 102 PB - Technischen Universität München CY - München AN - OPUS4-57744 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Ecke, Alexander T1 - The Fate of the Antibiotic Amoxicillin in the Aquatic Environment N2 - Contamination of the environment with antibiotics is of great concern as it promotes the evolution of antimicrobial resistances. In case of amoxicillin (AMX) in the aquatic environment, further risk arises from hydrolysis products (HPs) which can cause allergy. To assess these risks, a comprehensive investigation and understanding of the degradation of AMX is necessary. We investigated the hydrolysis rate of AMX in different types of water as well as the influence of temperature and irradiation. The content of the heavy metal ions copper and zinc was found to be crucial for the hydrolysis rate of AMX and stability of HPs. Eventually, a new degradation pathway for AMX could be elaborated and confirmed by tandem mass spectrometry (LC-MS/MS). T2 - Berliner Chemie in Praxis Symposium CY - Berlin, Germany DA - 07.10.2022 KW - Hydrolysis KW - Amoxicillin KW - LC-MS/MS KW - ICP-MS PY - 2022 AN - OPUS4-56026 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Insights at the point-of-need via immunoanalytical methods N2 - A vast number of emerging pollutants is being detected in the environment. Another lingering problem are health-threatening contaminants, such as mycotoxins, that deteriorate food and feed, and the pathogens themselves. Analytical methods, suitable for trace analysis, are needed that are desirably also fast, inexpensive and, if possible, robust and portable. It is set out, how immunoana-lytical, i.e., antibody-based methods, which are elaborated in a broad range of formats, can be profitably used to gain insights on the distribution and concentration trends of the target analytes at the point-of-need. T2 - Kolloquium der Fakultät für Pharmazie der Universität Porto CY - Porto, Portugal DA - 01.07.2022 KW - Immunoassay KW - Sensor KW - Antibody PY - 2022 AN - OPUS4-56781 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Schneider, Rudolf T1 - Advanced materials broadening the scope of antibody-based analytical methods N2 - Immunoanalytical Techniques, i.e., antibody-based analytical methods, have been used for decades in clinical diagnostics. What makes them attractive for other fields of application is their short time-to-result and high sensitivity. Microplate-based assays such as ELISA have been adopted early in environmental and food analysis. Yet, to make immunoassays even faster, more sensitive, robust, and, most desirable, portable, advanced materials, sometimes developed for other purposes, can be profitably used to achieve these goals. Materials can be novel labels, e.g., chemical or particle labels, such as fluorophores or nanoparticles. Carrier particles, such as magnetic or polymer beads, make it possible to adopt the assays to meso- or microfluidic set-ups and encoding them opens the path to multiplex analysis. Specialty electrodes can enable for higher sensitivity in electrochemical detection. All this broadens the scope of application and lowers effort and cost for analysis at the point-of-need. T2 - ChemForum - Kolloquium des Instituts für Strukturchemie CY - Lisbon, Portugal DA - 07.09.2022 KW - Immunoassay KW - Biosensor KW - Antibody PY - 2022 AN - OPUS4-56782 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -