title: Miniaturization of a transthyretin binding assay using a fluorescent probe for high throughput screening of thyroid hormone disruption in environmental samples.
abstract: Thyroid hormone (TH) disrupting compounds are potentially important environmental contaminants due to their possible adverse neurological and developmental effects on both humans and wildlife. Currently, the most successful bio-analytical method to detect and evaluate TH disruptors, which target the plasma transport of TH in environmental samples, is the radio-ligand thyroxine-transthyretin (T4-TTR) binding assay. Yet, costly materials and tedious handling procedures prevent the use of this assay in high throughput analysis that is nowadays urgently demanded in environmental quality assessment. For the first time a miniaturized fluorescence T4-TTR binding assay was developed in a 96 well microplate and tested with eight TH disrupting compounds. For most of the compounds, the sensitivity of the newly developed assay was slightly lower than the radio-ligand binding assay, however, throughput was enhanced at least 100-fold, while using much cheaper materials. The TH disrupting potency of 22 herring gull (Larus argentatus) egg extracts, collected from two different locations (Musvær and Reiaren) in Norway, was evaluated to demonstrate the applicability of the assay for environmental samples.
authors.affiliation: Department of Environment and Health, VU University Amsterdam, De Boelelaan 1108, 1081 HZ, Amsterdam, The Netherlands. Electronic address: xiyu.ouyang@vu.nl.
authors.forename: Xiyu
authors.initials: X
authors.lastname: Ouyang
authors.affiliation: Norwegian Institute for Water Research (NIVA), Gaustadalléen 21, N-0349, Oslo, Norway; Department of Chemistry, University of Oslo (UiO), PO Box 1033, Blindern, N-0316, Oslo, Norway.
authors.forename: Jean
authors.initials: J
authors.lastname: Froment
authors.affiliation: Department of Environment and Health, VU University Amsterdam, De Boelelaan 1108, 1081 HZ, Amsterdam, The Netherlands.
authors.forename: Pim E G
authors.initials: PEG
authors.lastname: Leonards
authors.affiliation: Akvaplan NIVA, FramCentre, N-9296, Tromsø, Norway.
authors.forename: Guttorm
authors.initials: G
authors.lastname: Christensen
authors.affiliation: Norwegian Institute for Water Research (NIVA), Gaustadalléen 21, N-0349, Oslo, Norway.
authors.forename: Knut-Erik
authors.initials: KE
authors.lastname: Tollefsen
authors.affiliation: Department of Environment and Health, VU University Amsterdam, De Boelelaan 1108, 1081 HZ, Amsterdam, The Netherlands.
authors.forename: Jacob
authors.initials: J
authors.lastname: de Boer
authors.affiliation: Norwegian Institute for Water Research (NIVA), Gaustadalléen 21, N-0349, Oslo, Norway.
authors.forename: Kevin V
authors.initials: KV
authors.lastname: Thomas
authors.affiliation: Department of Environment and Health, VU University Amsterdam, De Boelelaan 1108, 1081 HZ, Amsterdam, The Netherlands.
authors.forename: Marja H
authors.initials: MH
authors.lastname: Lamoree
mesh_terms: D000818:Animals; D001681:Biological Assay; D046091:Charadriiformes; D005456:Fluorescent Dyes; D008904:Miniaturization; D009664:Norway; D011228:Prealbumin / Q000378:metabolism*; D013974:Thyroxine / Q000378:metabolism*; D015053:Zygote
chemical_list: D005456:Fluorescent Dyes; D011228:Prealbumin; D013974:Thyroxine
keywords: Competitive binding assay; Fluorescein probe; Herring gull eggs; High throughput; Miniaturization; Thyroid hormone transport protein; Thyroxine