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Improved strategies for selection and characterization of new monoclonal anticarbamazepine antibodies during the screening process using feces and fluorescence polarization immunoassay

  • 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 newImmunoassays 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 mg L� 1 within 10 min using a high-throughput microtiter plate-based FPIA, and between 1.4 and 79 mg L� 1 within 5 min applying an automated cuvette-based FPIA instrument, and from 0.05–36 mg 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.zeige mehrzeige weniger

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Metadaten
Autoren/innen:Lidia Oberleitner, L.-A. Garbe, Ursula Dahmen-Levison, Rudolf J. Schneider
Dokumenttyp:Zeitschriftenartikel
Veröffentlichungsform:Verlagsliteratur
Sprache:Englisch
Titel des übergeordneten Werkes (Englisch):Analytical Methods
Jahr der Erstveröffentlichung:2016
Organisationseinheit der BAM:1 Analytische Chemie; Referenzmaterialien
1 Analytische Chemie; Referenzmaterialien / 1.8 Umweltanalytik
Veröffentlichende Institution:Bundesanstalt für Materialforschung und -prüfung (BAM)
Verlag:Royal Society of Chemistry
Verlagsort:London
Jahrgang/Band:8
Erste Seite:6883
Letzte Seite:6894
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Sanitär- und Kommunaltechnik; Umwelttechnik
Freie Schlagwörter:Antikörper; Arzneistoffe; Monoklonal; Pharmazeutika; Schadstoffe
Assay; Emerging Pollutants; Pesticide parathion-methyl; Pharmaceuticals; Quantification; Samples; Waste-water
Themenfelder/Aktivitätsfelder der BAM:Umwelt
Umwelt / Umweltschadstoffe
Analytical Sciences
Analytical Sciences / Spurenanalytik und chemische Zusammensetzung
DOI:https://doi.org/10.1039/c6ay01968d
URN:urn:nbn:de:kobv:b43-392473
URL:http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=Alerting&SrcApp=Alerting&DestApp=WOS&DestLinkType=FullRecord;UT=WOS:000385380800008
ISSN:1759-9660
Verfügbarkeit des Volltexts:Volltext-PDF für die Öffentlichkeit verfügbar ("Open Access")
Lizenz (Deutsch):License LogoCreative Commons - Namensnennung
Datum der Freischaltung:27.02.2017
Referierte Publikation:Ja
Datum der Eintragung als referierte Publikation:27.02.2017
Schriftenreihen ohne Nummerierung:Wissenschaftliche Artikel der BAM