A receptor-based chemiluminescence enzyme linked immunosorbent assay for determination of tetracyclines in milk
A receptor-based chemiluminescence enzyme linked immunosorbent assay for determination of tetracyclines in milk
复制标题
基于受体的化学发光酶联免疫吸附测定牛奶中的四环素类药物
DOI：
10.1016/j.ab.2018.10.017
复制
发表时间：
2019-01-01
影响因子：
2.9
通讯作者：
Wang, Jian Ping
中科院分区：
文献类型：
--
作者：
Wang, Ge;Zhang, Hui Cai;Wang, Jian Ping
该研究首次报道了基于受体的化学发光免疫分析法用于四环素类药物的分析。实验中将TetR蛋白基因转化Rosetta-gami(DE3)表达受体，并利用分子对接技术研究了其对5种四环素类药物的识别机制。结果表明，疏水作用和Pi-Pi键是TetR与四环素类药物结合的主要分子间作用力，D环是四环素类药物分子中主要的受体结合位置。然后以该受体为识别试剂，建立了直接竞争性酶联免疫吸附法测定牛奶中的5种药物，并用4-(咪唑-1-基)-苯酚增强鲁米诺-H2O2系统诱导光信号。牛奶中5种药物的检出限为5-16 pg/mL，标准强化空白奶的回收率为71.7%-95.8%。因此，该方法可作为一种简单、快速、超灵敏的工具来监测牛奶中四环素类药物的残留量。
This study for the first time reported a receptor based chemiluminescence immunoassay for analysis of tetracyclines. During the experiments, the gene of TetR protein was transformed into Rosetta-gami(DE3) to express a receptor, and its recognition mechanism for 5 tetracyclines was studied by using molecular docking technique. Results showed that hydrophobic interaction and Pi-Pi bond were the main intermolecular forces responsible for TetR-tetracyclines binding, and the D ring was the main receptor binding position in tetracyclines molecules. Then the receptor was used as recognition reagent to develop a direct competitive enzyme linked immunosorbent assay for determination of the 5 drugs in milk, and the light signal was induced with 4-(imidazol-1-yl)-phenol enhanced luminol-H2O2 system. The limits of detection for the 5 drugs in milk were in the range of 5-16 pg/mL, and the recoveries from the standards fortified blank milk were in the range of 71.7%-95.8%. Therefore, this method could be used as a simple, rapid, and ultra-sensitive tool to monitor the residues of tetracyclines in milk.