Institute of Pesticide and Environmental Toxicology, Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Zhejiang University, Hangzhou 310058, China.
Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Hangzhou 311400, China.
Biosensors (Basel). 2022 Sep 3;12(9):716. doi: 10.3390/bios12090716.
Residues of neonicotinoid pesticides have potential risks to food, environmental and biological safety. In this study, the hapten toward imidacloprid was adopted to gain antibodies. After molecular modeling, the electrostatic potentials of eight commonly-used neonicotinoid pesticides were individually calculated to analyze the structural similarity. Two representative compounds (imidacloprid and acetamiprid) with moderate similarity were rationally selected for hybridoma screening. Using this strategy, four clones of broad-specific monoclonal antibodies (mAbs) against multiple neonicotinoids were obtained, and the clone 6F11 exhibited the broadest spectrum to six neonicotinoid pesticides and two metabolites, with half-maximal inhibitory concentrations (IC) ranging from 0.20 to 5.92 ng/mL. Then, the novel antibody gene was sequenced and successfully expressed in full-length IgG form using mammalian cells. Based on the sensitive recombinant antibody, a gold lateral-flow immunosensing strip assay was developed and it was qualified for rapid detection of imidacloprid, clothianidin or imidaclothiz residues in food samples.
新烟碱类农药残留对食品、环境和生物安全具有潜在风险。本研究采用针对吡虫啉的半抗原获得抗体。经过分子建模，分别计算了 8 种常用新烟碱类农药的静电势，以分析其结构相似性。选择具有中等相似性的两种代表性化合物（吡虫啉和噻虫胺）进行杂交瘤筛选。使用该策略，获得了针对多种新烟碱类农药的 4 株广谱单克隆抗体（mAb）克隆，克隆 6F11 对 6 种新烟碱类农药和 2 种代谢物表现出最广的谱，半抑制浓度（IC）范围为 0.20-5.92ng/mL。然后，对新型抗体基因进行测序，并成功在哺乳动物细胞中以全长 IgG 形式表达。基于灵敏的重组抗体，开发了金侧向流动免疫传感条检测方法，可快速检测食品样品中的吡虫啉、噻虫啉或噻虫胺残留。
