Instituto Interuniversitario de Investigación en Bioingeniería y Tecnología Orientada al Ser Humano, Universitat Politècnica de València, Camino de Vera s/n, Edificio 8B-N, 46022 Valencia, Spain.
Food Chem. 2014 Jan 15;143:205-13. doi: 10.1016/j.foodchem.2013.07.121. Epub 2013 Aug 3.
Tetraconazole is currently used as a fungicide in fruit and vegetables. The aim of this work was the development of immunochemical techniques based on recombinant antibodies for the screening of tetraconazole residues in fruit juices. Recombinant antibodies were produced from a hybridoma cell line secreting a monoclonal antibody specific for tetraconazole and from lymphocytes of mice hyperimmunised with tetraconazole haptens conjugated to bovine serum albumin. From these antibodies, enzyme-linked immunosorbent assays in the conjugate-coated format were developed, which were able to detect tetraconazole standards down to 1ng/mL. From recovery studies with spiked samples, these immunoassays determined tetraconazole in orange and apple juices with acceptable reproducibility (coefficients of variation below 25%) and recoveries (ranging from 78% to 145%) for a screening technique. The analytical performance of RAb-based immunoassays was fairly similar to that of the MAb-based immunoassays. Due to their simplicity and high sample throughput, the developed recombinant-based immunoassays can be valuable analytical tools for the screening of tetraconazole residues in fruit juices at regulatory levels.
四氮唑目前被用作水果和蔬菜中的杀菌剂。本工作旨在开发基于重组抗体的免疫化学技术，用于筛选果汁中的四氮唑残留。从分泌针对四氮唑的单克隆抗体的杂交瘤细胞系和用与牛血清白蛋白偶联的四氮唑半抗原高度免疫的小鼠淋巴细胞中产生了重组抗体。从这些抗体中，开发了缀合涂层格式的酶联免疫吸附测定法，该方法能够检测到低至 1ng/mL 的四氮唑标准品。通过用加标样品进行回收研究，这些免疫测定法以可接受的重现性（变异系数低于 25%）和回收率（范围为 78%至 145%）确定了橙汁和苹果汁中的四氮唑，适用于筛选技术。基于 RAb 的免疫分析的分析性能与基于 MAb 的免疫分析相当。由于其简单性和高通量样品处理能力，开发的基于重组的免疫分析可以成为在监管水平上筛选果汁中四氮唑残留的有价值的分析工具。
