A novel monoclonal antibody （MAb） against oxytetracycline （OTC） was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay （ELISA）-based detection system. An OTC...A novel monoclonal antibody （MAb） against oxytetracycline （OTC） was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay （ELISA）-based detection system. An OTC-bovine serum albumin （BSA） conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2-4F, 7-3G, and 11-11A. An indirect competitive ELISA （icELISA） was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration （IC50） and percentage of cross-reactivity, respectively. MAb 2-4F exhibited the highest sensitivity, with an ICs0 of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross- reactivity to other unrelated antibiotics. When MAb 2-4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%-118% for an intra-assay and 96%-113% for an inter-assay. The coefficients of variation of the assays were 3.9%-13.9% and 5.5%-14.9%, respectively.展开更多
基金supported by the Ratchadaphiseksomphot Endowment Fund of Chulalongkorn University(No.RD_50-53_61)the National Research University Development Project(No.AM1023A)the Office of the Higher Education Commission,Thailand
文摘A novel monoclonal antibody （MAb） against oxytetracycline （OTC） was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay （ELISA）-based detection system. An OTC-bovine serum albumin （BSA） conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2-4F, 7-3G, and 11-11A. An indirect competitive ELISA （icELISA） was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration （IC50） and percentage of cross-reactivity, respectively. MAb 2-4F exhibited the highest sensitivity, with an ICs0 of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross- reactivity to other unrelated antibiotics. When MAb 2-4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%-118% for an intra-assay and 96%-113% for an inter-assay. The coefficients of variation of the assays were 3.9%-13.9% and 5.5%-14.9%, respectively.
