title: Development and validation of a sensitive monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay for the determination of the aflatoxin M1 levels in milk.
abstract: A sensitive monoclonal antibody (mAb) against aflatoxin M1 (AFM1) was generated to quickly monitor the AFM1 residues in milk. Then, a mAb-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established that utilizes simple sample preparation and clean-up methods. The obtained 3D8 mAb, which is an IgG1 isotype mAb, displayed an IC50 value of 64.75 ng L(-1) for AFM1 and did not exhibit measurable cross-reactivity with other aflatoxins and antibiotics. The decision limit (CCα, α = 1%), detection capability (CCβ, β = 5%), and LOQ value for the AFM1 matrix calibration method were 24 ng L(-1), 27.5 ng L(-1), and 35 ng L(-1) in the milk matrices, respectively. The AFM1 recovery ranged from 85.3% to 107.6%. The CVs were less than 13.8%. A positive correlation (r > 0.99) was observed between the ic-ELISA and HPLC-MS/MS results. This ic-ELISA would be a useful tool for screening the AFM1 residues in milk.
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
authors: Dapeng
authors: D
authors: Peng
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
authors: Bijia
authors: B
authors: Yang
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
authors: Yuanhu
authors: Y
authors: Pan
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
authors: Yulian
authors: Y
authors: Wang
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
authors: Dongmei
authors: D
authors: Chen
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China.
authors: Zhenli
authors: Z
authors: Liu
authors: Hubei Provincial Center for Disease Control and Prevention, Wuhan, Hubei 430079, China.
authors: Wenxiang
authors: W
authors: Yang
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China. Electronic address: tyf@mail.hzau.edu.cn.
authors: Yanfei
authors: Y
authors: Tao
authors: National Reference Laboratory of Veterinary Drug Residues (HZAU) and MAO Key Laboratory for Detection of Veterinary Drug Residues, Hubei Collaborative Innovation Center for Animal Nutrition and Feed Safety, Huazhong Agricultural University, Wuhan, Hubei 430070, China. Electronic address: yuan5802@mail.hzau.edu.cn.
authors: Zonghui
authors: Z
authors: Yuan
mesh_terms: D016607:Aflatoxin M1 / Q000737:chemistry*; D000818:Animals; D000911:Antibodies, Monoclonal; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007074:Immunoglobulin G; D051379:Mice; D008892:Milk / Q000737:chemistry*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D016607:Aflatoxin M1
keywords: Aflatoxin M(1); Indirect competitive enzyme-linked immunosorbent assay; Milk; Monoclonal antibody