抗磺胺对甲氧嘧啶单克隆抗体的研制及其特性分析

Preparation and characterization of monoclonal antibodies against sulfameter

目的制备针对磺胺对甲氧嘧啶的单克隆抗体,建立对该物质的免疫学检测方法。方法以BSA-磺胺对甲氧嘧啶为免疫原,免疫BALB/c小鼠,取脾细胞与小鼠Sp-2/0骨髓瘤细胞融合后,经筛选和亚克隆,建立杂交瘤细胞株。结果获得2株能稳定分泌抗磺胺对甲氧嘧啶抗体的细胞株。对抗体进行了特性分析,抗体的效价分别为1∶400 000和1∶1 630 000,抗体类型及亚类都为IgG1。其中,单克隆抗体1H10的亲和力为1.4×109L/mol,利用该抗体采用竞争间接ELISA法检测磺胺对甲氧嘧啶的范围是1025～16μg/mL,最低检测浓度是8μg/mL。单抗1H10与其他6种磺胺药(SMM、SMZ、SM2、SD、Sulfaquinoxaline Sodium、Sulfametopyrazine)无交叉反应。结论单克隆抗体1H10可用于研制免疫学方法检测磺胺对甲氧嘧啶残留的产品。

Objective To prepare monoclonal antibodies against sulfameter （SMD） and establish immunological detec- tion methods for these antibodies. Method BALB/c mice were immunized with BSA-SMD as the antigen, and the immunized spleen cells were fused with murine Sp2/0 myeloma cells. After subcloning and screening, the hybridomas secreting stably McAbs were established. Result Two hybridomas with high antibody titers were obtained. Their titers were 1 ： 400 000 and 1 ： 1 630 000 respectively, and the Ig subclasses were IgG1. The affinity constants （Kaffi） of IH10 was 1.4 × 109 L/mol. SMD range detected by the competitive indirect ELISA with McAb 1H10 was 1025 μg/mL to 16 μg/mL, with 8 μg/mL of the minimum detectable concentration. No cross-reactivity was seen between the antibody 1HIO and the other 6 sulfonamides. Conclusion The antibody 1H10 can be used in developing the immunoassay kits for the detection of SMD.