Mari Ghulam Mujtaba, Li Hongfang, Dong Baolei, Yang Huijuan, Talpur Aisha, Mi Jiafei, Guo Liuchuan, Yu Xuezhi, Ke Yuebin, Han Diangang, Wang Zhanhui
College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food, Beijing Laboratory for Food Quality and Safety, Beijing 100193, People's Republic of China.
Department of Genetic Toxicology, Shenzhen Center for Disease Control and Prevention, Shenzhen 518020, People's Republic of China.
Food Chem. 2021 Sep 1;355:129598. doi: 10.1016/j.foodchem.2021.129598. Epub 2021 Mar 17.
Derivatization is usually employed in immunoassay for detection of metabolites of nitrofurans and avoiding derivatization could be preferable to achieve an efficient screening. In the study, we designed four haptens of 4-hydroxybenhydrazide (HBH), the nifuroxazide metabolite. The effect of hapten structures on antibody affinity were evaluated and one monoclonal antibody was produced by using the Hapten C with a linear alkalane spacer arm. After optimization, an enzyme linked-immunosorbent assay (ELISA) was established with an 50% inhibition concentration of 0.25 ng mL for HBH, which could ensure the direct detection of HBH without derivatization. The limit of detection of the ELISA for HBH was 0.12 µg kg with the recoveries of 90.1-96.2% and coefficient of variation (CV) values lower than 9.1%. In conclusion, we produced several high affinity antibodies to HBH with new designed hapten and developed an icELISA for the direct detection of HBH without derivatization in chicken.
衍生化通常用于免疫分析中检测硝基呋喃的代谢物，而避免衍生化可能更有利于实现高效筛选。在本研究中，我们设计了四种4-羟基苯肼（HBH）（硝呋齐特代谢物）的半抗原。评估了半抗原结构对抗体亲和力的影响，并使用带有线性链烷间隔臂的半抗原C制备了一种单克隆抗体。经过优化，建立了一种酶联免疫吸附测定（ELISA）方法，其对HBH的50%抑制浓度为0.25 ng/mL，这可以确保直接检测HBH而无需衍生化。该ELISA对HBH的检测限为0.12 μg/kg，回收率为90.1 - 96.2%，变异系数（CV）值低于9.1%。总之，我们用新设计的半抗原制备了几种对HBH具有高亲和力的抗体，并开发了一种间接竞争ELISA用于直接检测鸡肉中未衍生化的HBH。
