TY - JOUR A1 - Oberleitner, Lidia A1 - Dahmen-Levison, U. A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Application of fluorescence polarization immunoassay for determination of carbamazepine in wastewater N2 - Carbamazepine is an antiepileptic drug that can be used as a marker for the cleaning efficiency of wastewater treatment plants. Here, we present the optimization of a fast and easy on-site measurement system based on fluorescence polarization immunoassay and the successful application to wastewater. A new monoclonal highly specific anti-carbamazepine antibody was applied. The automated assay procedure takes 16 min and does not require sample preparation besides filtration. The recovery rates for carbamazepine in wastewater samples were between 60.8 and 104% with good intra- and inter-assay coefficients of variations (less than 15 and 10%, respectively). This automated assay enables for the onsite measurement of carbamazepine in wastewater treatment plants. KW - On-site measurement KW - Wastewater treatment plant KW - Fluorescence polarization immunoassay KW - Assay optimization KW - Mix-and-read assay PY - 2017 U6 - https://doi.org/10.1016/j.jenvman.2017.01.063 SN - 0301-4797 SN - 1095-8630 VL - 193 SP - 92 EP - 97 PB - Elsevier Ltd. AN - OPUS4-39152 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Oberleitner, Lidia A1 - Eremin, S.A. A1 - Lehmann, Andreas A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Fluorescence polarization immunoassays for carbamazepine - comparison of tracers and formats N2 - For the antiepileptic drug and anthropogenic marker carbamazepine (CBZ), a fast and cost-effective immunoassay based on fluorescence polarization (FPIA) was developed. The required fluorophore conjugates were synthesized from different fluorescein and CBZ derivatives. The most suitable tracer was CBZ–triglycine–5-(aminoacetamido)fluorescein. Additionally, the applicability of the assay in tubes and on microtiter plates was tested. The first format can be performed in a portable instrument and therefore can be applied in field measurements. The measurement of an individual sample can be carried out within 4 min. This assay shows a measurement range of 2.5–1000 µg L-1 and a test midpoint (or IC50) of 36 µg L-1. The FPIA performed on microtiter plates is useful for the assay development and is suitable for a very high throughput (up to 24 samples in 20 min). The test midpoint of this assay is 13 µg L-1 and the measurement range is 1.5–300 µg L-1. Furthermore, this assay requires smaller sample volumes and less reagents, including the crucial amount of antibody. The applicability of both assays to spiked surface water samples was evaluated. The recovery rates vary between 66–110% on microtiter plates and 81–140% in tubes. PY - 2015 U6 - https://doi.org/10.1039/c5ay00617a SN - 1759-9660 SN - 1759-9679 VL - 7 IS - 14 SP - 5854 EP - 5861 PB - RSC Publ. CY - Cambridge AN - OPUS4-33821 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Oberleitner, Lidia A1 - Grandke, Julia A1 - Mallwitz, F. A1 - Resch-Genger, Ute A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Fluorescence polarization immunoassays for the quantification of caffeine in beverages N2 - Homogeneous fluorescence polarization immunoassays (FPIAs) were developed and compared for the determination of caffeine in beverages and cosmetics. FPIAs were performed in cuvettes in a spectrometer for kinetic FP measurements as well as in microtiter plates (MTPs) on a multimode reader. Both FPIAs showed measurement ranges in the µg/L range and were performed within 2 and 20 min, respectively. For the application on real samples, high coefficients of variations (CVs) were observed for the performance in MTPs; the CVs for FPIAs in cuvettes were below 4%. The correlations between this method and reference methods were satisfying. The sensitivity was sufficient for all tested samples including decaffeinated coffee without preconcentration steps. The FPIA in cuvettes allows a fast, precise, and automated quantitative analysis of caffeine in consumer products, whereas FPIAs in MTPs are suitable for semiquantitative high-throughput screenings. Moreover, specific quality criteria for heterogeneous assays were applied to homogeneous immunoassays. KW - caffeine KW - Decaffeinated coffee KW - Fluorescence polarization immunoassay KW - Homogeneous immunoassay KW - Quality assurance criteria PY - 2014 U6 - https://doi.org/10.1021/jf4053226 SN - 0021-8561 SN - 1520-5118 VL - 62 IS - 11 SP - 2337 EP - 2343 PB - American Chemical Society CY - Columbus, Ohio AN - OPUS4-30613 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Oberleitner, Lidia A1 - Garbe, L.-A. A1 - Dahmen-Levison, Ursula A1 - Schneider, Rudolf T1 - Improved strategies for selection and characterization of new monoclonal anticarbamazepine antibodies during the screening process using feces and fluorescence polarization immunoassay N2 - Immunoassays are suitable tools for high-throughput screenings. The prerequisite for accurate determinations by these methods is the selection of an excellent antibody. The production and selection of monoclonal antibodies is usually a tedious process. In this study, new strategies for improving antibody production and characterization were applied. This includes the monitoring of the immunization progress in mice through antibodies extracted from feces, which allows a time-resolved and animal-friendly monitoring of the immune response. Additionally, fluorescence polarization immunoassay (FPIA) could be successfully applied for fast and easy examination of cell culture supernatants and the investigation of antibody/antigen interactions including kinetics and fluorescence properties. These methods simplify the selection of the optimal antibody. As a target analyte, carbamazepine was chosen. This is a widely used antiepileptic drug which also frequently occurs in the environment. The new antibody enables CBZ determination in the concentration range 0.66–110 µg L-1 within 10 min using a high-throughput microtiter plate-based FPIA, and between 1.4 and 79 µg L-1 within 5 min applying an automated cuvette-based FPIA instrument, and from 0.05–36 µg L-1 using ELISA. The measurements were performed at a non-equilibrium state which improved the sensitivity and selectivity of the assays. Due to low cross-reactivity especially towards the main CBZ metabolite and other pharmaceuticals (<1%), this antibody gives the opportunity for application in medical and environmental analyses. KW - Antikörper KW - Emerging Pollutants KW - Schadstoffe KW - Monoklonal KW - Pharmazeutika KW - Arzneistoffe KW - Assay KW - Pesticide parathion-methyl KW - Waste-water KW - Samples KW - Pharmaceuticals KW - Quantification PY - 2016 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:b43-392473 SN - 1759-9660 VL - 8 SP - 6883 EP - 6894 PB - Royal Society of Chemistry CY - London AN - OPUS4-39247 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Grandke, Julia A1 - Oberleitner, Lidia A1 - Resch-Genger, Ute A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Quality assurance in immunoassay performance - carbamazepine immunoassay format evaluation and application on surface and waste water N2 - Carbamazepine (CBZ) is one of the most frequently detected pharmaceuticals in water samples. For the determination of this anthropogenic marker, various immunoassay formats were tested and evaluated in order to identify the most suitable one. For these direct competitive assays, the analyte was labelled with the enzyme horseradish peroxidase (HRP) or alkaline phosphatase (AP), and seven substrates with specific detection properties were used. The quality criteria for the standard curves were fulfilled by all HRP assays and the chemiluminescence AP format. Furthermore, intra- and inter-plate coefficients of variation as a measure of the achievable precision were determined for the samples. The application of the AP assays to surface water was unfeasible due to CBZ concentrations below the quantifiable concentration range. Surface as well as waste water samples could be analyzed with the HRP assays. Here, the HRP assay employing the chromogenic substrate 3,3',5,5'-tetramethylbenzidine yielded the best results. PY - 2013 U6 - https://doi.org/10.1039/c3ay40702k SN - 1759-9660 SN - 1759-9679 VL - 5 IS - 15 SP - 3754 EP - 3760 PB - RSC Publ. CY - Cambridge AN - OPUS4-28860 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - JOUR A1 - Grandke, Julia A1 - Oberleitner, Lidia A1 - Resch-Genger, Ute A1 - Garbe, L.-A. A1 - Schneider, Rudolf T1 - Quality assurance in immunoassay performance - comparison of different enzyme immunoassays for the determination of caffeine in consumer products N2 - Enzyme immunoassays with optical detection are amongst the most widely used bioanalytical tools. We defined seven parameters for the quality assessment of immunoassays that were addressed in a systematic study of direct and indirect immunoassays, using the enzymes horseradish peroxidase (HRP) and alkaline phosphatase (AP), the chromogenic substrates 3,3',5,5'-tetramethylbenzidine (TMB) and para-nitrophenyl phosphate, and the fluorescent substrates 3-(4-hydroxyphenyl)propionic acid and 4-methylumbelliferyl phosphate. The same monoclonal antibody against caffeine was used throughout the study. The four quality parameters regarding the standard curve were the test midpoint (sensitivity), the measurement range, the relative dynamic range of the signal, and the goodness of fit of the adjusted four-parameter logistic function. All HRP immunoassays showed a higher sensitivity compared to the AP assays. On the basis of all four criteria, it was established that the direct assay format is superior to the indirect format, the immunoassay using HRP TMB fulfilling all requirements best. In a second step, caffeine concentrations in 24 beverage and cosmetics samples were determined and three more quality parameters were assessed with this application. The direct HRP TMB assay showed one of the best intra- and inter-plate precisions and the best accuracy, defined by the correlation of results with those from the chosen reference method liquid chromatography tandem mass spectrometry (LC-MS/MS). Considering all criteria, HRP TMB seems to be the enzyme substrate system of choice preferably used in the direct assay format. KW - Enzyme immunoassay KW - Caffeine KW - Enzyme substrate KW - Competitive assay KW - Assay format KW - Direct KW - Indirect PY - 2013 U6 - https://doi.org/10.1007/s00216-012-6596-0 SN - 1618-2642 SN - 1618-2650 VL - 405 IS - 5 SP - 1601 EP - 1611 PB - Springer CY - Berlin AN - OPUS4-27639 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -