TY - CONF A1 - Ecke, Alexander A1 - Schneider, Rudolf T1 - Immunanalytische Bestimmung von β-Lactam-Antibiotika und deren Hydrolyseprodukten im Trinkwasser N2 - Die speziellen Herausforderungen der immunanalytischen Bestimmung von β-Lactam-Antibiotika in (Trink-)Wasserproben werden anhand eines neu entwickelten ELISA (enzyme-linked immunosorbent assay) für das Breitbandantibiotikum Amoxicillin diskutiert. Es konnte gezeigt werden, dass die Hydrolyse des Analyten einen positiven Einfluss auf die Erkennung durch den Antikörper hat. Das entsprechende Hydrolyseprodukt, zu dem der Antikörper folglich eine höhere Affinität aufweist, wurde identifiziert und ein entsprechendes Verhalten wurde auch für andere β-Lactam-Antibiotika aus der Gruppe der Penicilline beobachtet. Eine Strategie für die Analyse von Wasserproben hinsichtlich Kontamination mit den hier betrachteten β-Lactamen und deren Hydrolyseprodukten wurde ausgearbeitet. T2 - Wasser 2020 - Jahrestagung der Wasserchemischen Gesellschaft CY - Meeting was canceled DA - 18.05.2020 KW - ELISA KW - Immunoassay KW - Antibiotika KW - Hydrolyseprodukte KW - Amoxicillin PY - 2020 UR - https://www.wasserchemische-gesellschaft.de/images/pdf_veranstaltungen/2020/Wasserchemische_Gesellschaft_Tagungsband_Wasser_2020_s.pdf SP - 466 EP - 470 AN - OPUS4-51061 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Silva, C.P. A1 - Lima, D.L.D. A1 - Schneider, Rudolf A1 - Otero, M. A1 - Esteves, V.I. T1 - Evaluation of the anthropogenic input of caffeine in surface waters of the north and center of Portugal by ELISA N2 - This study comprises the development of an enzyme-linked immunosorbent assay (ELISA) for the quantification of caffeine in complex aqueous matrices without any sample clean-up procedure. Salinity and dissolved organic matter were selected as potential interfering agents. The addition of a sample buffer containing bovine serum albumin (BSA) prior to the sample was found to decrease the influence of those interfering agents. The working range of the developed method was 0.1–100 µg L- 1. Quantification of caffeine was possible in 43 out of 51 real aqueous samples, at values between < LOD and 15 µg L- 1. Results correlate well with those obtained by LC–MS/MS. To the best of our knowledge this is the first study dealing with the quantification of caffeine in Portugal's surface waters. KW - Caffeine KW - Surface water KW - ELISA KW - Anthropogenic markers KW - Koffein KW - Eintrag KW - Oberflächenwasser KW - Gewässer KW - Abwasser KW - Antikörper KW - Immunoassay PY - 2014 DO - https://doi.org/10.1016/j.scitotenv.2014.01.120 SN - 0048-9697 VL - 479-480 SP - 227 EP - 232 PB - Elsevier B.V. CY - Amsterdam AN - OPUS4-30450 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Baldofski, Stefanie A1 - Hoffmann, Holger A1 - Lehmann, Andreas A1 - Breitfeld, Stefan A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Enzyme-linked immunosorbent assay (ELISA) for the anthropogenic marker isolithocholic acid in water N2 - Bile acids are promising chemical markers to assess the pollution of water samples with fecal material. This study describes the optimization and validation of a direct competitive enzyme-linked immunosorbent assay for the bile acid isolithocholic acid (ILA). The quantification range of the optimized Assay was between 0.09 and 15 mg/L. The assay was applied to environmental water samples. Most studies until now were focused on bile acid fractions in the particulate phase of water samples. In order to avoid tedious sample preparation, we undertook to evaluate the dynamics and significance of ILA levels in the aqueous phase. Very low concentrations in tap and surface water samples made a pre-concentration step necessary for this matrix as well as for wastewater treatment plant (WWTP) effluent. Mean recoveries for spiked water samples were between 97% and 109% for tap water and WWTP influent samples and between 102% and 136% for WWTP effluent samples. 90th percentiles of intra-plate and inter-plate coefficients of variation were below 10% for influents and below 20% for effluents and surface water. ILA concentrations were quantified in the range of 33-72 mg/L in influent, 21-49 ng/L in effluent and 18-48 ng/L in surface water samples. During wastewater Treatment the ILA levels were reduced by more than 99%. ILA concentrations of influents determined by ELISA and LC-MS/MS were in good agreement. However, findings in LC-ELISA experiments suggest that the true ILA levels in concentrated samples are lower due to interfering effects of matrix compounds and/or cross-reactants. Yet, the ELISA will be a valuable tool for the performance check and comparison of WWTPs and the localization of fecal matter input into surface waters. KW - ELISA KW - LC-MS/MS KW - Bile acids KW - Fecal marker KW - Surface water KW - Wastewater PY - 2016 DO - https://doi.org/10.1016/j.jenvman.2016.08.023 SN - 0301-4797 VL - 182 SP - 612 EP - 619 AN - OPUS4-37668 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Carvalho, J.J.D. A1 - Bahlmann, Arnold A1 - Schneider, Rudolf T1 - ELISA goes Berlin - Bestimmung von Carbamazepin, Cetrizin und Coffein in Berliner Gewässern N2 - Sensitive und selektive Immunoassays unterstützen Untersuchungen im Vollzug der EU Wasserrahmenrichtlinie. Sie erlauben, eine hohe Anzahl von Proben ohne Anreicherungsschritte zu untersuchen und ermöglichen daher den Aufbau eines zeitlich und räumlich engmaschigen Messnetzes. Mit Hilfe von ELISA (enzymelinked immunosorbent assay) wurde ein Gewässerscreening in Berlin durchgeführt. KW - ELISA KW - Koffein KW - Immunoassay KW - Oberflächenwasser KW - Wasseranalytik PY - 2012 SN - 0016-3538 VL - 4 SP - 278 EP - 281 PB - GIT-Verlag CY - Darmstadt AN - OPUS4-27518 LA - deu 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 - Figueira, E. A1 - Freitas, R. T1 - Effects of single and combined exposure of pharmaceutical drugs (carbamazepine and cetirizine) and a metal (cadmium) on the biochemical responses of R. philippinarum N2 - In the aquatic environment, organisms are exposed to complex mixtures of contaminants which may alter the toxicity profile of each compound, compared to its toxicity alone. Pharmaceutical drugs (e.g. carbamazepine (CBZ) and cetirizine (CTZ)) and metals (e.g. cadmium (Cd)) are among those contaminants that co-occur in the environment. However, most studies concerning their toxicity towards aquatic species are based on single exposure experiments. Thus, the present study aimed to evaluate single and combined effects of Cd and CBZ or CTZ (single conditions: Cd, CTZ, CBZ; combined conditions: CTZ+Cd, CBZ+Cd) on biomarkers related to oxidative stress and energy metabolism in the edible clam Ruditapes philippinarum, by exposing the organisms for 28 days to environmentally relevant concentrations of these contaminants. The biomarkers studied were: i) the electron transport system activity, protein and glycogen contents (indicators of organisms’ metabolic status and energy reserves); ii) lipid peroxidation and the ratio between reduced and oxidized glutathione (indicators of oxidative stress); iii) superoxide dismutase and catalase activities (enzymes indicators of antioxidant defence) and iv) activity of glutathione S-transferases (family of enzymes indicators of biotransformation capacity). Results obtained showed that the uptake of Cd and CBZ was not affected by the combined presence of the contaminants. However, for CTZ, the uptake was higher in the presence than in the absence of Cd. Concerning toxicity data, in general, the combined exposures (CTZ+Cd, CBZ+Cd) had lower biological effects than the contaminants alone. Nevertheless, our data showed that despite the low concentrations tested, they were enough to exert biological effects that differed between single and combined treatments, evidencing the need to conduct more co-exposure studies to increase the environmental relevance of the gathered data. KW - Biomarker KW - Arzneimittel KW - Metalle KW - Invertebraten KW - ELISA KW - Carbamazepine KW - Cetirizine PY - 2018 DO - https://doi.org/10.1016/j.aquatox.2018.02.011 SN - 0166-445X SN - 1879-1514 VL - 198 SP - 10 EP - 19 PB - Elsevier CY - Amsterdam, NL AN - OPUS4-44902 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Almeida, Â. A1 - Freitas, R. A1 - Calisto, V. A1 - Esteves, V. I. A1 - Schneider, Rudolf A1 - Soares, A. M. V. M. A1 - Figueira, E. A1 - Campos, B. A1 - Barata, C. T1 - Effects of carbamazepine and cetirizine under an ocean acidification scenario on the biochemical and transcriptome responses of the clam Ruditapes philippinarum N2 - Several works evaluated the toxicity of pharmaceutical drugs and climate related changes in invertebrates but few explored the combined effects of both stressors, namely considering their mode of action (MoA). Carbamazepine (CBZ) and cetirizine (CTZ) are pharmaceutical drugs detected in the environment and the toxicity derived from the combined effects of these drugs with ocean acidification (OA) is poorly explored. Thus, the present study investigated the biochemical parameters related to an oxidative stress response and the transcription of genes related to the MoA of CBZ (1.0 mg/L) and CTZ (0.6 mg/L) in the clam Ruditapes philippinarum chronically exposed (28 days) to control (7.8) and low (7.5) pH conditions. The results obtained showed that despite the clams accumulated both drugs, at low pH the clams exposed to CTZ decreased drug concentration and BCF values (CTZ uptake: 2.0 ± 0.5 ng/g fresh weight; BCF: 3.8 ± 0.9) in comparison with clams exposed to control pH (CTZ uptake: 2.9 ± 0.3 ng/g fresh weight; BCF: 5.5 ± 0.6). No oxidative stress was induced by the exposure to CBZ or CTZ at each pH level, but the transcription of several genes related with the MoA (neurotransmission, immunity and biomineralization) was altered by low pH, drug exposure and the combination of both stressors. At both pH conditions, CBZ increased the transcription of GABA receptor gene (neurotransmission) and CTZ led to a decrease of Perlucin gene (biomineralization) transcription. The transcription of MyD88 gene (immunity) decreased at low pH (7.5) combined with drug exposure (CBZ or CTZ). Thus, it was highlighted that the interaction of drug exposure and low pH conditions can change bivalves’ sensitivity to drugs or alter drugs toxicity. KW - Carbamazepine KW - Biomarker KW - ELISA KW - Biochemische Parameter PY - 2018 DO - https://doi.org/10.1016/j.envpol.2017.12.121 SN - 0269-7491 VL - 235 SP - 857 EP - 868 PB - Elsevier Ltd. CY - Amsterdam, NL AN - OPUS4-44739 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Lima, D.L.D. A1 - Silva, C.P. A1 - Schneider, Rudolf A1 - Esteves, V.I. T1 - Development of an ELISA procedure to study sorption of atrazine onto a sewage sludge-amended luvisol soil N2 - Pesticides may contaminate ground and surface waters and one of the major factors governing this property is soil sorption. Sorption can be assessed by batch equilibrium technique which produces lots of extracts with high dissolved organic carbon concentration in which the pesticide concentration has to be determined. We developed an ELISA procedure to analyse atrazine based on polyclonal antibodies (C193) for which tracer structure and dilutions of immunochemical reagents were adapted to fit the purpose. After a 1000-fold dilution (or after an SPE clean-up procedure) extracts of a sewage-sludge amended luvisol (used as an example application of the methodology developed) could be reliably analysed. The Freundlich model is able to describe adsorption for this system (r² = 0.977) delivering a distribution coefficient KF of 1.6 ± 0.2 (mg kg-1) (mg L-1)-N and an isotherm nonlinearity factor N of 0.70 ± 0.09. KW - ELISA KW - Atrazine KW - Sorption PY - 2011 DO - https://doi.org/10.1016/j.talanta.2011.06.024 SN - 0039-9140 VL - 85 IS - 3 SP - 1494 EP - 1499 PB - Elsevier B.V. CY - Amsterdam AN - OPUS4-24305 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Braeutigam, P. A1 - Franke, M. A1 - Schneider, Rudolf A1 - Lehmann, Andreas A1 - Stolle, A. A1 - Ondruschka, B. T1 - Degradation of carbamazepine in environmentally relevant concentrations in water by hydrodynamic-acoustic-cavitation (HAC) N2 - The antiepileptic drug carbamazepine is one of the most abundant pharmaceuticals in the German aquatic environment. The effect of low carbamazepine concentrations (1–50 µg L-1) is discussed controversially, but ecotoxicological studies revealed reproduction toxicity, decreased enzymatic activity and bioaccumulation in different test organisms. Therefore, as a preventive step, an efficient and cost-effective technique for wastewater treatment plants is needed to stop the entry of pharmaceuticals into the aquatic environment. Cavitation, the formation, growth, and subsequent collapse of gas- or vapor-filled bubbles in fluids, was applied to solve this problem. The technique of Hydrodynamic-Acoustic-Cavitation was used showing high synergistic effect. Under optimized conditions carbamazepine (5 µg L-1) was transformed by pseudo-first order kinetics to an extent of >96% within 15 min (27% by hydrodynamic cavitation, 33% by acoustic cavitation). A synergistic effect of 63% based on the sum of the single methods was calculated. Carbamazepine concentrations were monitored by a sensitive and selective immunoassay and after 60 min no known metabolites were detectable by LC-MS/MS. KW - Carbamazepine KW - Degradation KW - Hydrodynamic cavitation KW - Acoustic cavitation KW - ELISA PY - 2012 DO - https://doi.org/10.1016/j.watres.2012.02.013 SN - 0043-1354 VL - 46 IS - 7 SP - 2469 EP - 2477 PB - Elsevier CY - Amsterdam AN - OPUS4-25735 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Schneider, Rudolf A1 - Carvalho, A. M. A1 - Manicardi, A. A1 - Montes, C. V. A1 - Gunnoo, S. B. A1 - Madder, A. T1 - Decoration of trastuzumab with short oligonucleotides: synthesis and detailed characterization N2 - Trastuzumab (Herceptin) is an FDA-approved therapeutic antibody currently employed in the treatment of metastatic stages of breast cancer. Herein, we propose a simple, fast and cost-effective methodology to conjugate trastuzumab with 22-mer 5' thiol-modified oligonucleotides using a bifunctional crosslinker. The conjugates were successfully characterized by MALDI-ToF MS and SDS-PAGE, obviating the need for enzymatic digestion and difficult chromatographic separations. Furthermore, ELISA was performed to ensure that trastuzumab activity is not affected by oligonucleotide conjugation. KW - MALDI-ToF KW - ELISA PY - 2017 DO - https://doi.org/10.1039/c7ob02216f SN - 1477-0539 VL - 15 IS - 42 SP - 8923 EP - 8928 AN - OPUS4-43328 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Kuhne, Maren A1 - Dippong, Martin A1 - Flemig, Sabine A1 - Hoffmann, Katrin A1 - Petsch, K. A1 - Schenk, J.A. A1 - Kunte, Hans-Jörg A1 - Schneider, Rudolf T1 - Comparative characterization of mAb producing hapten-specific hybridoma cells by flow cytometric analysis and ELISA N2 - A novel method that optimizes the screening for antibody-secreting hapten-specific hybridoma cells by using flow cytometry is described. Cell clones specific for five different haptens were analyzed. We selectively double stained and analyzed fixed hybridoma cells with fluorophore-labeled haptens to demonstrate the target-selectivity, and with a fluorophore-labeled anti-mouse IgG antibody to characterize the level of surface expression of membrane-bound IgGs. ELISA measurements with the supernatants of the individual hybridoma clones revealed that antibodies from those cells, which showed the highest fluorescence intensities in the flow cytometric analysis, also displayed the highest affinities for the target antigens. The fluorescence intensity of antibody-producing cells corresponded well with the produced antibodies' affinities toward their respective antigens. Immunohistochemical staining verified the successful double labeling of the cells. Our method makes it possible to perform a high-throughput screening for hybridoma cells, which have both an adequate IgG production rate and a high target affinity. KW - Immunization KW - Hapten KW - Monoclonal antibodies KW - Hybridoma KW - Flow cytometry KW - ELISA KW - Estradiol KW - Estrone KW - Digoxigenin KW - Zearalenone KW - Aflatoxin KW - CLSM PY - 2014 DO - https://doi.org/10.1016/j.jim.2014.07.004 SN - 0022-1759 SN - 1872-7905 VL - 413 SP - 45 EP - 56 PB - Elsevier B.V. CY - Amsterdam AN - OPUS4-32322 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -