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Der Vortrag beschreibt den Stand der Arbeiten an der BAM, Fachbereich Umweltanalytik zur Entwicklung von Sensoren für die Wasseranalytik. Er stellt die bisherigen Arbeiten da, die ausgehend von ELISAs, zunehmend portable Plattformen wie FPIA und LFIA umfassen. Beas-based Formate werden von mikrotiterplatten-basierten Techniken auf mikrofluidische Lab-on-Chip-Systeme übertragen, die bei geeigneter Miniaturisierung und Automatisierung als Sensorsysteme angesehen werden können. Der Vortrag endet mit einem Blick auf ein laufendes Monitoring-Projekt von anthropogenen Markern in Abwasser und charakterisiert die Treiber, die die Entwicklung von Sensoren für die Wasseranalytik in den letzten Jahren vorantreiben.
The environmental fate of the frequently used broad-spectrum β-lactam antibiotic amoxicillin (AMX) is of high concern regarding the potential evolution of antimicrobial resistance (AMR). Moreover, it is known that AMX is prone to hydrolysis, yielding a variety of hydrolysis products (HPs) with yet unknown effects. Studies to identify those HPs and investigate their formation mechanisms have been reported but a long-term study on their stability in real water samples was missing. In this regard, we investigated the hydrolysis of AMX at two concentration levels in four distinct water types under three different storage conditions over two months. Concentrations of AMX and four relevant HPs were monitored by an LC-MS/MS method revealing pronounced differences in the hydrolysis rate of AMX in tap water and mineral water on the one hand (fast) and surface water on the other(slow). In this context, the occurrence, relative intensities, and stability of certain HPs are more dependent on the water type than on the storage condition. As clarified by ICP-MS, the main difference between the water types was the content of the metals copper and zinc which are supposed to catalyze AMX hydrolysis demonstrating an effective method to degrade AMX at ambient conditions.
Functionalized Ti3C2Tx nanosheets based biosensor for point-of-care detection of SARS-CoV‑2 antigen
(2023)
MXenes are considered a promising class of two-dimensional materials with extraordinary physical and electrochemical properties. Distinguished features like high specific surface area and outstanding electrical conductivity make them suitable for electrochemical biosensing applications. Here, we report the development of a biosensor involving the functionalized MXene−titanium carbide nanosheets (Ti3C2Tx-NS) and monoclonal antibodies against the SARS-CoV-2 nucleocapsid protein (anti-SARS-CoV-2 mAb) to design a point-of-care device for detection of the SARS-CoV-2 nucleocapsid protein (SARS-CoV-2 NP) antigen. Few-layered titanium carbide nanosheets (denoted as FL-Ti3C2Tx-NS) have been synthesized using a single-step etching and delamination method and characterized using optical and electron microscopy techniques revealing the suitability for immunosensing applications. Binding studies revealed the excellent affinity between the biosensor and the SARS-CoV-2 NP. Electrochemical detection of SARS-CoV-2 NP is performed using differential pulse voltammetry and read by a smartphone-based user interface. The proposed FL-Ti3C2Tx-NS based biosensor offers the detection of SARS-CoV-2 NP with a limit of detection of 0.91 nM in a wide detection range in spiked saliva samples. Additionally, there is no cross-reactivity in the presence of potential interferants like SARS-CoV-2 spike glycoprotein and bovine serum albumin. These findings demonstrate the potential of MXenes in developing a rapid and reliable tool for SARS-CoV-2 NP detection. While we report the biosensing of SARS-CoV-2 NP, our system also paves the way for the detection of other SARS-CoV-2 antigens like spike protein or other biomolecules based on antigen−antibody interactions.
Im Vorhaben „MARKERIA - Bestimmung von anthropogenen Markerkonzentrationen für die SARS-CoV-2 Quantifizierung mittels eines Hochdurchsatzverfahrens (ELISA)“ geht es um die Bestimmung von Carbamazepin, Diclofenac, Koffein und einer endogenen Gallensäure als potentielle Marker für die Kalibrierung der SARS-CoV-2 Quantifizierung im Abwasser. Der Vortrag wurde gehalten im Rahmen der 22. Sitzung des Begleitkreises des übergeordneten Projektes ESI-CorA. Er gab den Projektstand wieder, den die BAM bis dahin erzielt hatte. Die Marker waren in sämtlichen untersuchten Abwasser-Zulaufproben enthalten und zeigten spezifische Trends. Weitere Korrelationen müsse die weitergehende Datenauswertung ergeben.A
Immunoassays, based on analyte recognition and capture by highly selective antibodies with high affinity, are intensively used in all fields of laboratory diagnostics and in screen-ings of food and environmental samples. Yet, for many purposes, online sensors are desir-able, and, in principle, all immunoassay tech-niques can be integrated into lab-on-chip set-ups that can work as continuous monitoring devices. Yet, the challenge remains to devel-op platforms and elements that are fit for a quick transition of laboratory microplate as-says to immunosensors.
The poster describes how molecular biology, especially recombinant expression of proteins, in this case, an enzyme, can underpin developments of biosensors. The fumonisin oxidase produced by the fungus Aspergillus niger (AnFAO) is highly selective for the toxic mycotoxin fumonisin. Its structure and sequence has been published before. We took this information and expressed the enzyme in E. coli. The enzyme proved active and could be employed in an amperometric biosensor for the detection of the mycotoxin.
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.
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.