Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany.
Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany; Institute of Biotechnology, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Cau Giay, Ha Noi, Viet Nam.
Sci Total Environ. 2017 Dec 15;605-606:507-513. doi: 10.1016/j.scitotenv.2017.06.077. Epub 2017 Jun 30.
Arxula adeninivorans-based yeast cell assays for the detection of steroid hormones demonstrated their efficiency for the determination of total hormone activity in a variety of samples using a microtiter plate format. In this study, a preliminary chromatographic separation using thin-layer chromatography plates is introduced in order to allow a rapid identification of the compounds responsible for this hormonal activity. The yeast whole cell assay can then be performed on the plate, producing a detectable signal where a steroid hormone is present. Simultaneous detection of estrogens, progestogens and androgens on the same plate in the picogram range was achieved, while keeping the assay as simple and affordable as possible. The assay requires a single incubation of the thin-layer chromatography plate and the detection of reporter protein production can be performed by fluorescence scanning of the plate at different wavelengths. The chromatographic separation allows the separation of several estrogens, androgens and progestogens, thus making its application for 'real world' samples very useful. In this work, different water-based samples from environmental origins were used to demonstrate the capacity of this new bioassay. Trials showed that most samples, with the exception of complex samples such as wastewater influent, can be assayed.
基于 Arxula adeninivorans 的酵母细胞检测法可用于检测甾体激素，其采用微孔板形式，可用于各种样品中总激素活性的测定，证明了其效率。在本研究中，引入了初步的薄层色谱分离，以允许快速鉴定出导致这种激素活性的化合物。然后可以在平板上进行酵母全细胞检测，如果存在甾体激素，则会产生可检测的信号。同时可以在同一平板上检测到皮克级范围内的雌激素、孕激素和雄激素，同时保持检测尽可能简单和经济实惠。该检测法只需对薄层色谱板进行一次孵育，并且可以通过在不同波长下扫描平板来检测报告蛋白的产生。该色谱分离可分离多种雌激素、雄激素和孕激素，因此非常适用于“实际”样品。在这项工作中，使用了来自环境来源的不同水样来证明这种新生物测定法的能力。试验表明，除了废水进水等复杂样品外，大多数样品都可以进行检测。
