论文部分内容阅读
目的 制备抗弓形虫主要表膜P30抗原单克隆抗体 (McAb)并进行鉴定 ,为弓形虫病的诊断、抗原的提纯及亚单位疫苗的研制等提供可靠依据。方法 用RH株弓形虫速殖子膜抗原为免疫原免疫BALB/C小鼠 ,取其脾细胞与小鼠SP2 / 0骨髓瘤细胞融合 ,筛选出能够稳定分泌高滴度抗P30抗原McAb杂交瘤细胞株 ,并测定McAb免疫球蛋白亚类和单抗效价 ,用IFAT进行单抗识别的抗原定位 ,并通过SDS PAGE和Western blot分析鉴定。结果 本实验获得了 2株抗P30抗原的杂交瘤细胞株E3 和G2 ,分泌的抗体滴度分别为 1∶2 5 880和 1∶10 2 6 0 ,其分泌的抗体与肺孢子虫、隐孢子虫等抗原均不发生交叉反应 ,2株单抗均属IgG1亚类 ,且识别的抗原定位于速殖子表膜。结论 制备的抗P30抗原的杂交瘤细胞株能分泌高滴度和特异性的单克隆抗体
Objective To prepare and identify McAbs against P30 antigen of Toxoplasma gondii and to provide a reliable basis for the diagnosis of toxoplasmosis, the purification of antigen and the development of subunit vaccines. Methods BALB / C mice were immunized with Toxoplasma gondii RH antigen of RH strain. The spleen cells were fused with mouse SP2 / 0 myeloma cells, and the McAbs that could secrete high titer anti-P30 McAb The McAb immunoglobulin subclasses and monoclonal antibody titer were determined, and the antigens recognized by the monoclonal antibody were localized by IFAT and identified by SDS PAGE and Western blot analysis. Results Two hybridoma cell lines E3 and G2 with anti-P30 antigen were obtained in this experiment. The secreted antibody titers were 1:25 5880 and 1:10 2 6 0, respectively. The secreted antibodies were expressed against Pneumocystis carinii, Cryptosporidium Insects and other antigens are not cross-reactive, two monoclonal antibodies are IgG1 subclass, and the identified antigen located in the tachyzoite surface membrane. Conclusion The prepared anti-P30 antigen hybridoma cell line can secrete high titers and specific monoclonal antibodies