白喉类毒素单克隆抗体的制备及初步鉴定

Preparation and identification of monoclonal antibodies against dophtheriae toxoid

目的:制备并鉴定白喉毒素单克隆抗体杂交瘤细胞株,为白喉的快速检测及致病机制的研究提供有力的抗体工具。方法:用白喉类毒素做抗原免疫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白喉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株持续分泌抗白喉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可以和白喉类毒素发生特异性反应。结论:成功制备了抗白喉类毒素单克隆抗体,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 a hybridoma cell line secreting monoclonal antibody（McAb） against diphtheriae toxoid to develop a rapid assay for tetanus and to investigate the pathogenesis of the virus.Methods：BALB/c mouse was immunized by purified diphtheriae toxoid.Hybridoma cell lines secreting monoclonal antibodies（McAbs） against diphtheriae 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 monoclonal antibodies specifically against diphtheriae toxoid were obtained,and respectively named as 1E12,1E11 and 1G10.Indirect EL ISA and Western-blot assay showed that the McAbs 1E12,1E11 and 1G10 could specifically recognize diphtheriae toxoid.Conclusion：The specific anti-diphtheriae toxoid McAbs are developed and these McAbs may be useful in the development of detection assay and basic research of the pathogenesis of tetanus.