破伤风类毒素单克隆抗体的制备及初步鉴定

Preparation and Identification of Monoclonal Antibodies Against Tetanic Toxoid

目的:制备并鉴定破伤风类毒素单克隆抗体杂交瘤细胞株,为破伤风的快速检测及致病机制的研究提供实验材料。方法:用破伤风类毒素做抗原免疫BALB/C小鼠,取其脾细胞与小鼠骨髓瘤细胞融合,获得分泌高滴度针对破伤风类毒素的杂交瘤细胞株,测定单抗免疫球蛋白亚类及单抗效价,用间接ELISA和Western-blot检测单克隆细胞株的特异性。结果:通过细胞融合和克隆化,筛选出3株持续分泌抗破伤风类毒素单克隆抗体的杂交瘤细胞株1E12、1E11、1G10。间接ELISA和Western-blot检测结果表明1E12、1E11、1G10可以和破伤风类毒素发生特异性反应。结论:成功制备了抗破伤风类毒素单克隆抗体,为制备免疫诊断试剂盒和抗体药物的开发奠定了基础。

Objective： To establish and prepare hybridoma cell line secreting monoclonal antibody （McAb） against tetanic toxoid, to develop a rapid assay for Tetanus and investigate the pathogenesis of the virus. Methods： BALB/c mouse was immunized by purified tetanic toxoid. Hybridoma cell lines secreting monoclonal antibodies （McAbs） against tetanic toxoid were screened after the fusion of mouse splenic cells with SP2/0 cells. The Ig subtypes and titer of the McAbs were assayed with EL ISA and Western-blot assay. Results： After cell fusion and subcloning, three hybridoma cell lines secreting monoelonal antibodies specifically against tetanic toxoid were obtained, and respectively named as 1E12, 1E11 and 1G10. Indirect EL ISA and Western-blot assay showed that the McAbs 1E12, 1E11and 1G10 could specifically recognize tetanic toxoid. Conclusion： The specific anti-tetanic toxoid McAbs are developed and these McAbs may be useful in the development of detection assay and basic research of the pathogenesis of Tetanus.