Development of a monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for screening of diethylstilbestrol in animal-derived foods
Bofei Fu,
Haijiao Gao,
Cuilan Fang,
Guyue Cheng,
Hui Wang,
Yulian Wang,
Haihong Hao,
Xu Wang,
Lingli Huang,
Dapeng Peng
Affiliations
Bofei Fu
Jiulongpo Center for Disease Control and Prevention, Chongqing, 400039, China
Haijiao Gao
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China; Fengdu Animal Disease Prevention and Control Center, Chongqing, 408200, China
Cuilan Fang
Jiulongpo Center for Disease Control and Prevention, Chongqing, 400039, China; Corresponding author.
Guyue Cheng
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China; Corresponding author.
Hui Wang
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China
Yulian Wang
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China
Haihong Hao
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China
Xu Wang
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China
Lingli Huang
National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China
Dapeng Peng
Center for Veterinary Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, China; National Reference Laboratory of Veterinary Drug Residues (HZAU) and MOA Key Laboratory for the Detection of Veterinary Drug Residues, Huazhong Agricultural University, Wuhan, Hubei, 430070, China; Corresponding author. Center for Veterinary Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, China.
Diethylstilbestrol (DES), a synthetic non-steroid estrogen, it has been prohibited from being added to animal feed for any purposes. Herein, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a specific monoclonal antibody (mAb) was developed for the rapid screening of DES. Primarily, conjugates of mono-O-3-carboxypropyl diethylstilbestrol with keyhole limpet hemocyanin were used to raise a specific mAb, 1B7, which had IC50 value for DES of 213.0 ng L−1. The limits of detection and limits of quantification value for DES in animal-derived foods ranged from 68.1 to 103.1 ng L−1 and 100.8–192.7 ng L−1, respectively. The DES recovery ranged from 70.1 % to 103.1 %, with coefficients of variation below 13.9 %. A positive correlation (R2 = 0.997) was observed between the results of ic-ELISA and HPLC-MS/MS for milk. In order to inspect its detection effect, milk and animal-derived foods were chosen as the testing object. The results showed that this ic-ELISA method (specific mAb, 1B7) can effectively examine for DES residues.