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

作者：Peng, Dapeng; Yang, Bijia; Pan, Yuanhu; Wang, Yulian; Chen, Dongmei; Liu, Zhenli; Yang, Wenxiang; Tao, Yanfei^*; Yuan, Zonghui^*

来源：Toxicon, 2016, 113: 18-24.

DOI：10.1016/j.toxicon.2016.02.006

• Aflatoxin M-1
• Monoclonal antibody
• Indirect competitive enzyme-linked immunosorbent assay
• Milk

A sensitive monoclonal antibody (mAb) against aflatoxin M-1 (AFM(1)) 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 (CC alpha, alpha = 1%), detection capability (CC beta, beta = 5%), and LOQ value for the AFM(1) matrix calibration method were 24 ng L-1, 27.5 ng L-1, and 35 ng L-1 in the milk matrices, respectively. The AFM(1) 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 AFM(1) residues in milk.

• 出版日期2016-4
• 单位华中农业大学; 湖北省疾病预防控制中心