申请人:The European Union,
represented by the European Commission
公开号:EP3064946A1
公开(公告)日:2016-09-07
Uses of ethyl 5,11-dihydroindolo[3,2-b]carbazole-6-carboxylate as a fluorescent ligand probe, preferably in a binding assay for quantitative analysis in combination with a recombinant aryl hydrocarbon receptor (AhR) protein. A method for detection or quantitative analysis of suspected aryl hydrocarbon receptor (AhR) ligands in a sample, the method comprising the steps of: (a) providing a sample possibly containing at least one known or unknown AhR ligand; (b) mixing said sample with a composition comprising a recombinant AhR protein bound to ethyl 5,11-dihydroindolo[3,2-b]carbazole-6-carboxylate; and (c) determining the presence or the total amount of said at least one known or unknown AhR ligand in the sample by fluorescence spectroscopy. Use e.g. for screening of unknown compounds to determine their binding ability to AhR; for analyzing unknown chemical mixtures to assess the presence of ligands able to bind to AhR; for screening of Polycyclic Aromatic Hydrocarbon (PAHs) from water, food or sediment; for screening of PAHs or dioxin-like compounds in water, food or sediment; for testing for pollutant removal in water and waste water treatment plants; for testing for water reuse; for testing the presence of dioxin-like compounds in incinerator plants; or for testing of bodily fluids of humans or animals exposed to PAHs or dioxin-like compounds.
5,11-二氢吲哚并[3,2-b]咔唑-6-羧酸乙酯作为荧光配体探针的用途,优选与重组芳基烃受体(AhR)蛋白结合进行定量分析的结合测定。一种检测或定量分析样品中可疑芳基烃受体(AhR)配体的方法,该方法包括以下步骤(a) 提供可能含有至少一种已知或未知 AhR 配体的样品;(b) 将所述样品与包含与 5,11-二氢吲哚并[3,2-b]咔唑-6-羧酸乙酯结合的重组 AhR 蛋白的组合物混合;以及 (c) 通过荧光光谱测定样品中所述至少一种已知或未知 AhR 配体的存在或总量。例如筛选未知化合物,确定其与 AhR 的结合能力;分析未知化学混合物,评估是否存在能够与 AhR 结合的配体;筛选水、食物或沉积物中的多环芳香烃(PAHs);筛选水、食物或沉积物中的多环芳香烃或二恶英类化合物;用于检测水和废水处理厂的污染物去除情况;用于检测水的再利用情况;用于检测焚化炉中是否存在二恶英类化合物;或用于检测暴露于多环芳烃或二恶英类化合物的人类或动物的体液。