Institute of Nutrition, Department of Nutritional Toxicology, Friedrich Schiller University of Jena, Dornburger Strasse 24, Jena, Germany.
Free Radic Res. 2012 Mar;46(3):276-85. doi: 10.3109/10715762.2011.652627. Epub 2012 Feb 6.
3-Nitrotyrosine (3NT) is known as an important indicator of nitrosative stress and has been linked to various diseases. Our aim was to develop an indirect ELISA (enzyme-linked immunosorbent assay) method suitable for the detection of protein-bound 3NT in clinical plasma and serum samples. Nitrated protein standards and reduced protein standards were prepared. Limit of detection was determined for standards; recovery and reproducibility were determined for human plasma samples. The limit of detection for this method is 1.82±0.56 pmol/mg protein. Mean recovery of standards was 95%. 3NT concentration in plasma samples of obese and normal weight subjects was determined to be between 2 pmol/mg and 19 pmol/mg. No time-consuming sample preparation or expensive laboratory equipment is required, and applied antibodies are commercially available. Sensitivity, rapid analysis time, possibilities of high throughput applications and small sample volumes make this ELISA attractive for use in clinical laboratories.
3-硝基酪氨酸（3NT）是一种重要的硝化应激指标，与多种疾病有关。我们的目的是开发一种间接酶联免疫吸附测定（ELISA）方法，适用于检测临床血浆和血清样本中蛋白质结合的 3NT。制备了硝化蛋白标准品和还原蛋白标准品。测定了标准品的检测限；测定了人血浆样品的回收率和重现性。该方法的检测限为 1.82±0.56 pmol/mg 蛋白。标准品的平均回收率为 95%。肥胖和正常体重受试者的血浆样本中 3NT 的浓度在 2 pmol/mg 至 19 pmol/mg 之间。该方法不需要耗时的样品制备或昂贵的实验室设备，并且应用的抗体可商购获得。该 ELISA 具有灵敏度高、分析时间短、高通量应用的可能性和小样本量等优点，因此在临床实验室中具有吸引力。
