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Development of a sensitive Indirect Competitive Enzyme-Linked Immunosorbent Assay (ic-ELISA) based on the monoclonal antibody for the detection of the imidaclothiz residue
Authors:Fang Song  Zhang Bin  Ren Ke-Wei  Cao Meng-Meng  Shi Hai-Yan  Wang Ming-Hua
Institution:Department of Pesticide Science, College of Plant Protection, Nanjing Agricultural University , Jiangsu Key Laboratory of Pesticide Science, Key Laboratory of Monitoring and Management of Crop Diseases and Pest Insects, Ministry of Agriculture, Nanjing 210095, People's Republic of China.
Abstract:An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on monoclonal antibodies (MoAbs) for imidaclothiz was developed. The hapten of imidaclothiz was synthesized and conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to form the artificial antigens. MoAbs were obtained by immunizing BALB/c mice. Under the optimized conditions (10% methanol, 0.14 M Na(+), and pH 7.4), the half-maximal inhibition concentration (IC(50)) was 0.0875 ± 0.0034 mg/L and the limit of detection (IC(20)) was 0.0178 ± 0.0018 mg/L for imidaclothiz. There were no obvious cross-reactivities with most of the structural analogues of neonicotinoid insecticides, except imidacloprid. The recoveries of imidaclothiz in environmental and agricultural samples, including tap water, paddy water, soil, and cabbage, ranged from 80.43 to 113.83%, well within the requirements of residue detection. These results showed that this immunoassay could be used for the determination of imidaclothiz in environmental and agricultural samples.
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