Abstract

Mepanipyrim is a new-generation anilinopyrimidine pesticide that is employed nowadays to fight fungal diseases in a variety of crops. Rapid and user-friendly analytical methods that can complement the instrumental techniques are demanded. To analyse residues of this toxic compound in foodstuffs, monoclonal antibodies were generated for the first time, and an enzyme-linked immunosorbent assay was developed. In the present study, a series of haptens mimicking the mepanipyrim molecule, and with a linker at alternative sites of the molecular framework, were designed and prepared by total synthesis in order to study the influence of the linker tethering site over the binding properties of the generated antibodies. This strategy afforded high-affinity and specific monoclonal antibodies to mepanipyrim. Interestingly, the highest sensitivity was reached with antibodies that had been raised using haptens with the spacer arm at the most characteristic moiety of the analyte, the 1-propynyl group. Moreover, the influence of the triple bond was investigated both in immunizing bioconjugates and in assay antigens. Finally, a competitive enzyme-linked immunosorbent assay was optimized and characterized, showing an IC₅₀ value of 0.3 μg/L for mepanipyrim. Good accuracy and precision values were found when grape juice and wine samples were analysed.

中文翻译：

---

基于单克隆抗体的葡萄汁和葡萄酒中甲吡吡啶残留敏感性分析的免疫分析

抽象的

Mepanipyrim是新一代的苯胺基嘧啶杀虫剂，如今已用于对抗多种农作物中的真菌疾病。需要能够与仪器技术互补的快速且用户友好的分析方法。为了分析食品中这种有毒化合物的残留，首次产生了单克隆抗体，并开发了一种酶联免疫吸附测定法。在本研究中，通过全合成设计和制备了一系列模仿mepanipyrim分子的半抗原，并在分子框架的其他位点带有一个连接子，以研究该连接子束缚位点对它们的结合特性的影响。产生的抗体。该策略提供了针对甲吡嘧啶的高亲和力和特异性单克隆抗体。有趣的是，使用半抗原在间隔物臂的最特征部分为1-丙炔基的半抗原产生的抗体可达到最高的灵敏度。此外，在免疫生物缀合物和测定抗原中都研究了三键的影响。最后，对竞争性酶联免疫吸附测定进行了优化和表征，显示了IC₅₀的浓度为0.3μg/ L。分析葡萄汁和葡萄酒样品时，发现了良好的准确度和精密度值。