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目的 :制备抗磺胺嘧啶 (SD)的单克隆抗体 (mAb) ,并研制SD快速检测试剂盒。方法 :以SD BSA的偶联物作为抗原免疫BALB/c小鼠 ,采用杂交瘤技术制备抗SD的mAb。并用抗SDmAb和SD HRP酶标抗原 ,建立一种可检测样品中的游离SD分子的竞争法ELISA。结果 :获得 5株可分泌抗SDmAb的杂交瘤细胞 1A1、1B8、1E4、2C1和 3D9。其中 1A1、1B8和 1E4为IgG1,2C1和 3D9为IgG2。试剂盒的半数抑制率 (I5 0 )为 9.3μg/L ,理论最低检出量达到 0 .6μg/L ,对于不同样品中SD的回收率均高于 60 %。除与SD反应以外 ,该试剂盒对其他磺胺类药物检测的交叉反应均小于 3 %。结论 :成功地制备了抗SD的mAb ,并建立了一种可快速检测各种样品中SD的竞争ELISA试剂盒
Objective: To prepare anti-sulfadiazine (mAb) monoclonal antibody (mAb) and to develop SD rapid test kit. METHODS: BALB / c mice were immunized with the conjugate of SD BSA as antigen and the anti-SD mAb was prepared by hybridoma technique. And using anti-SDmAb and SD HRP enzyme-labeled antigens to establish a competitive ELISA that detects free SD molecules in the sample. Results: Five hybridomas secreting anti-SDmAb 1A1, 1B8, 1E4, 2C1 and 3D9 were obtained. Among them, 1A1, 1B8 and 1E4 were IgG1, 2C1 and 3D9 were IgG2. The half inhibition rate (I5 0) of the kit was 9.3 μg / L and the theoretical minimum detectable amount was 0.6 μg / L. The recoveries of SD in different samples were higher than 60%. In addition to the reaction with SD, the kit has less than 3% cross-reactivity to other sulfonamides. CONCLUSIONS: Anti-SD mAb was successfully prepared and a competitive ELISA kit was developed for the rapid detection of SD in various samples