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目的:制备抗多菌灵小鼠单克隆抗体(mAb),并初步鉴定其特性,为快速检测蘑菇、干菇类中残留的多菌灵提供基础。方法:将小分子的多菌灵交联在大分子载体匙孔嘁血蓝蛋白(KLH)和牛血清白蛋白(BSA)上,KLH交联物用于免疫BALB/c小鼠,采用B细胞杂交瘤技术,通过融合,筛选制备抗多菌灵mAb,并通过ELISA方法测定抗体亲和力,检测mAb的交叉反应,及初步竞争抑制检测。结果:通过ELISA筛选出7株稳定分泌特异性抗体的杂交瘤细胞株,其亚型均为IgG1。与同属于苯并咪唑类杀菌剂的苯菌灵和甲基硫菌灵没有交叉反应。竞争抑制检测结果显示MC-3细胞株竞争效果较好。结论:获得1株高特异性、高亲和力、能稳定分泌抗多菌灵抗体的杂交瘤细胞株,为进一步研发多菌灵残留免疫检测技术和产品奠定基础。
Objective: To prepare anti-carbendazim monoclonal antibody (mAb) and preliminary identification of its characteristics, providing a basis for the rapid detection of residual carbendazim in mushrooms and dried mushrooms. Methods: The small molecule carbendazim was crosslinked on KLH and bovine serum albumin (BSA). KLH conjugate was used to immunize BALB / c mice. B cell hybridization Tumor technique, anti-carbendazim mAb was prepared by fusion, screening, measuring antibody affinity by ELISA, detecting mAb cross-reaction, and preliminary competition inhibition test. Results: Seven hybridoma cell lines stably secreting specific antibodies were screened by ELISA, and the subtypes were all IgG1. With the same benzimidazole fungicides benomyl and thiophanate-methyl cross-reaction. Competition inhibition test results show that MC-3 cell line competition is better. Conclusion: Obtaining a hybridoma cell line with high specificity and high affinity, which can stably secrete anti-carbendazim antibody, which lays the foundation for further research on the residual immunoassay techniques and products of carbendazim.