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 DO - 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 - CONF A1 - Oberleitner, Lidia T1 - Fluorescence polarization immunoassay - the fast alternative to ELISA T2 - Schnell, schneller, Optik - wie optische Technologien die Lebensmittel- und Umweltanalytik optimieren CY - Berlin DA - 2014-11-27 PY - 2014 AN - OPUS4-32143 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberleitner, Lidia T1 - New monoclonal anti-carbamazepine antibody for application in fluorescence polarization immunoassays T2 - Analytica Conference 2014 CY - Munich, Germany DA - 2014-04-01 PY - 2014 AN - OPUS4-30516 LA - deu 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 UR - https://nbn-resolving.org/urn:nbn:de:kobv:b43-392473 DO - https://doi.org/10.1039/c6ay01968d 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 - 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 DO - 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 - CONF A1 - Oberleitner, Lidia T1 - Fluorescence polarization immunoassay - Fast Screening method for antibody characterization T2 - Tag der Biotechnologie CY - Berlin, Germany DA - 2015-07-16 PY - 2015 AN - OPUS4-34514 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberleitner, Lidia T1 - Fluorescense Polarization Immunoassays for Caffeine T2 - Euroanalysis 2013 CY - Warsaw, Poland DA - 2013-08-25 PY - 2013 AN - OPUS4-28995 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberleitner, Lidia T1 - Fluorescense Polarization Immunoassays for Carbamazepine T2 - 24th Symposium on Pharmaceutical and Biomedical Analysis CY - Bologna, Italy DA - 2013-06-30 PY - 2013 AN - OPUS4-28946 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberleitner, Lidia T1 - Application and evaluation of carbamazepine immunoassays T2 - ANAKON 2013 CY - Essen, Germany DA - 2013-03-04 PY - 2013 AN - OPUS4-27814 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Oberleitner, Lidia T1 - Comparison of Heterogeneous and homogeneous Immunoassays T2 - Trends in Diagnostics CY - Tübingen, Germany DA - 2013-10-07 PY - 2013 AN - OPUS4-29238 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -