抗磺胺嘧啶单克隆抗体的制备及其ELISA检测试剂盒的建立

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

Preparation of monoclonal antibody against sulfadiazine and development of an ELISA kit

AIM: To prepare monoclonal antibody (mAb) against sulfadiazine (SD) and develop an ELISA kit for rapidly detecting residual SD in different kinds of samples. METHODS: BALB/c mice were immunized with the conjugate of SD and BSA, and then anti-SD mAb was prepared by hybridoma technique. The purified ascitic mAb and HRP-labeled SD were used to establish a competitive ELISA for detection of SD in samples. RESULTS: 5 hybridoma cell lines secreting anti-SD mAbs 1A1, 1B8, 1E4, 2C1 and 3D9 were obtained. 1A1, 1B8 and 1E4 belonged to IgG1, while 2C1 and 3D9 belonged to IgG2. The I50 and theoretical minimum detectable amount of the kit was 9.3 microg/L and 0.6 microg/L, respectively. The recovery rates of the kit for SD in different kinds of samples were higher than 60%. The cross-reaction rate of the kit for other sulfanilamide drugs was lower than 3%. CONCLUSION: 5 mAbs against SD have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for rapidly detecting SD can meet the needs of detection of SD in different samples.