Development and validation of a sensitive monoclonal antibody-based ic-ELISA for the aflatoxin M1 in milk (Abstract) | ||
| Annals of Agricultural Science, Moshtohor | ||
| Article 16, Volume 56, 4th ICBAA - Serial Number 1, 2018, Pages 75-76 PDF (203.29 K) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/assjm.2018.57331 | ||
| Author | ||
| Dapeng Peng* | ||
| College of veterinary medicine, Huazhong Agricultural University, Wuhan, China | ||
| Abstract | ||
| Aflatoxin M1 (AFM1), a hydroxylated metabolite of aflatoxin B1 (AFB1), possesses the carcinogenic, mutagenic, and toxic activities. To protect the health of animals and humans, it is clear that the need for an effective residue-monitoring programme to detect the AFM1 residues. In this study, a modified procedure was used to prepare the hapten AFB1 and AFM1 derivatives. Then, the prepared antigen AFM1-CMO-KLH was used to inoculate female Balb/c mice to prepare a sensitive monoclonal antibody (mAb) against AFM1. After cell fusion and culture several times, the hybridoma cell line, 3D8, which was of the IgG1 isotype, was selected to obtain a highly sensitive mAb. The obtained 3D8 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. Based on this mAb, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was established that utilizes simple sample preparation and clean-up methods. 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. | ||
| Keywords | ||
| Monoclonal antibody (mAb); Aflatoxin M1; Milk | ||
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