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[目的]以新型抗病毒剂创制品种毒氟磷为研究对象,通过杂交瘤技术建立抗毒氟磷及其类似物的高特异性杂交瘤细胞系,应用体内诱生腹水制备单克隆抗体,开发出检测毒氟磷及其类似物的酶联免疫分析方法。[方法]利用2-甲酰苯氧乙酸、氨基酸合成半抗原,通过碳二亚胺法合成人工抗原,免疫动物并通过杂交瘤技术得到一株杂交瘤细胞株DHS3A2。[结果]在制备的单克隆抗体mAb-DHS3A2的基础上,通过对有机溶剂、pH值、离子强度等因素的筛选比较,优化了毒氟磷间接竞争ic-ELISA,方法的IC50值为(9.62±0.59)μg/L,检出限为(0.3±0.05)μg/L。[结论]该方法快速、准确,适用于不同基质中毒氟磷农药残留快速检测。
[Objective] To establish a high-specificity hybridoma cell line of antitoxic fluorine-phosphorus and its analogs by using new antiviral agent to produce drug-resistant flurbiprofen and to produce monoclonal antibody by in vivo induced ascites. Enzyme-linked immunosorbent assay for the detection of toxoflurane and its analogues. [Method] With the use of 2-formylphenoxyacetic acid and amino acid to synthesize hapten, the artificial antigen was synthesized by carbodiimide method, the animal was immunized and a hybridoma cell line DHS3A2 was obtained by hybridoma technology. [Result] Based on the monoclonal antibody mAb-DHS3A2 prepared, the indirect competition of icofloxacin ic-ELISA was optimized by screening and comparing the factors such as organic solvent, pH and ionic strength. The IC50 value was 9.62 ± 0.59) μg / L, the detection limit was (0.3 ± 0.05) μg / L. [Conclusion] The method was rapid and accurate and could be applied to the rapid detection of F and P pesticide residues in different substrates.