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Development of a monoclonal antibody-based immunoaffinity chromatography and a sensitive immunoassay for detection of spinosyn A in milk, fruits, and vegetables

Lan, Jianqiang, Zhao, Hongwei, Jin, Xiaotuo, Guan, Huai, Song, Yuan, Fan, Yongmei, Diao, Xiaoping, Wang, Baomin, Han, Qian
Food control 2019 v.95 pp. 196-205
crops, cross reaction, detection limit, enzyme-linked immunosorbent assay, fruits, haptens, high performance liquid chromatography, immunoaffinity chromatography, insect control, macrolides, milk, molecular models, monitoring, monoclonal antibodies, spinosad, vegetables
Spinosad has been extensively used for insect control in agricultural crops. In the present study, a novel succinic derivative of spinosyn A as the hapten was synthesized, and an indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed using a new monoclonal antibody (designed as mAb3E6) specifically against spinosyn A. A mAb3E6-based immunoaffinity column was developed for enrichment extraction of spinosyn A and reduction of matrix interference. The half maximum inhibitory concentration and limit of detection (LOD) of the icELISA were 4.11 ng/mL and 0.63 ng/mL, respectively. The icELISA had average fortified recoveries from 78.1% to 103.2% and the validation results correlated well with high-performance liquid chromatography. The mAb3E6 showed cross reactivity of 80.4%, 75.7% and less than 0.1% with spinosyns B, D and other macrolides, respectively. Interactions of the antibody and macrolides were further analyzed by molecular docking. The developed immunoaffinity chromatography-assisted icELISA is potentially a sensitive and convenient tool for monitoring spinosad residues in milk, fruits, and vegetables.