ABSTRACT

Van Lake is the third largest closed lake in the world and the biggest lake in Turkey. An ELISA method has developed with the aim of determining the pollution caused by estrogens and estrogen-like chemicals that have come to the lake Van in recent years. First, the vitellogenin in estrogen-treated male fish plasma was purified by ion exchange chromatography, injected into rats, and the obtained polyclonal antibodies were tested for specificity by Western blot and immunohistochemical methods. Immunohistochemical labeling of the vitellogenin-synthesized liver resulted in the intense marking of the liver of the animals injected with estrogen, while no markings were observed in the control group. The limit of detection of the developed enzyme-linked immunosorbent assay was 4.6 µg L⁻¹, and the working range was 7.8 to 2000 µg L⁻¹. Intra- and inter-assay variations were 13.0 % and 13.3%. The highest level of vitellogenin in male fishes measured was 23.56 µg mL⁻¹.

摘要

凡湖是世界第三大封闭湖，也是土耳其最大的湖泊。开发了一种 ELISA 方法，旨在确定近年来进入 Van 湖的雌激素和雌激素类化学物质造成的污染。首先，雌激素处理的雄鱼血浆中的卵黄蛋白通过离子交换层析纯化，注射到大鼠体内，得到的多克隆抗体通过蛋白质印迹和免疫组化方法检测特异性。卵黄蛋白原合成肝脏的免疫组织化学标记导致注射雌激素的动物肝脏出现强烈标记，而在对照组中未观察到任何标记。开发的酶联免疫吸附试验的检测限为 4.6 µg L ^-1，工作范围为 7.8 至 2000 µg L ^-1。测定内和测定间变化分别为 13.0% 和 13.3%。测得的雄性鱼卵黄蛋白原的最高水平为 23.56 µg mL ^-1。