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Determination of pharmacologically active compounds in wastewater by a bead-based flow-cytometric immunoarray

  • The assessment of water quality demands for multi-target compatible analytical methods. On the one hand, the influence of sewage on natural surface waters must be routinely monitored. On the other hand, estimation of removal efficiencies of pollutants, such as drug residues, is in the focus of industrial and public wastewater treatment. Immunoassays, such as ELISA, are often applied in this area, however, it is only possible to measure a single analyte in one measurement. In contrast, array technology is capable for measuring multiple substances in parallel. Here we present a four-plex bead-based flow cytometric assay for measuring three drugs (carbamazepine, an antiepileptic drug, diclofenac, an antiphlogistic and caffeine, a psychoactive substance) and the bile acid isolithocholic acid, which is proposed to be used as a fecal marker. In this suspension array core-shell particles, consisting of a polystyrene core and a silica shell, are used. They can be easily encoded with aThe assessment of water quality demands for multi-target compatible analytical methods. On the one hand, the influence of sewage on natural surface waters must be routinely monitored. On the other hand, estimation of removal efficiencies of pollutants, such as drug residues, is in the focus of industrial and public wastewater treatment. Immunoassays, such as ELISA, are often applied in this area, however, it is only possible to measure a single analyte in one measurement. In contrast, array technology is capable for measuring multiple substances in parallel. Here we present a four-plex bead-based flow cytometric assay for measuring three drugs (carbamazepine, an antiepileptic drug, diclofenac, an antiphlogistic and caffeine, a psychoactive substance) and the bile acid isolithocholic acid, which is proposed to be used as a fecal marker. In this suspension array core-shell particles, consisting of a polystyrene core and a silica shell, are used. They can be easily encoded with a fluorophore which is introduced to the core while the shell is functionalized with amino groups to which the analytes or their derivates are bound via NHS chemistry. They are the competitor binding sites for the antibodies. For the determination of the pollutants, the beads are incubated with a mixture of the analyte-specific antibodies. The molecules on the surface of the beads and the free analyte in solution compete for the binding sites of the antibodies. Bead-bound antibodies can then be visualized via dye-labelled secondary antibodies. For read-out a flow cytometer is used to virtually separate the differently encoded beads. In order to obtain highly selective binding of the antibodies, we investigated different types of surface modifications to overcome unspecific binding, finding that a PEG-based surface is suitable to support our immunoassay format. The resulting multiplexing assay is appropriate to detect the marker substances in the low µg/L range.zeige mehrzeige weniger

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Metadaten
Autoren/innen:Peter Carl, Rudolf J. Schneider
Koautoren/innen:Dominik Sarma, Knut Rurack
Dokumenttyp:Vortrag
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2017
Organisationseinheit der BAM:1 Analytische Chemie; Referenzmaterialien
1 Analytische Chemie; Referenzmaterialien / 1.8 Umweltanalytik
1 Analytische Chemie; Referenzmaterialien / 1.9 Chemische und optische Sensorik
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Sanitär- und Kommunaltechnik; Umwelttechnik
Freie Schlagwörter:Anthropogenic markers; Bead-based assay; Flow-cytometry; Immunoassay
Themenfelder/Aktivitätsfelder der BAM:Umwelt
Umwelt / Umweltschadstoffe
Analytical Sciences
Analytical Sciences / Sensorik
Veranstaltung:Pittcon 2017
Veranstaltungsort:Chicago, IL, USA
Beginndatum der Veranstaltung:05.03.2017
Enddatum der Veranstaltung:09.03.2017
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:24.03.2017
Referierte Publikation:Nein