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为制备抗弓形虫主要表膜抗原 P30单克隆抗体并进行鉴定 ,进而为弓形虫病诊断、抗原的提纯及亚单位疫苗的制备等提供可靠依据。用弓形虫速殖子膜蛋白为免疫原免疫 BAL B/ c小鼠 ,取其脾细胞与小鼠骨髓瘤细胞 SP2 / 0融合 ,筛选出能够稳定分泌高滴度抗 P30单克隆抗体的杂交瘤细胞株 ,并测定单克隆抗体免疫球蛋白亚类和抗体效价 ,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。结果获得了两株抗 P30抗原的杂交瘤细胞株 E3和 G2 ,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 Ig G1亚类 ,且识别的抗原定位于速殖子表膜。结果表明 ,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。
In order to prepare and identify the monoclonal antibodies against P30, the major surface antigens of Toxoplasma gondii, and to provide a reliable basis for the diagnosis of toxoplasmosis, the purification of antigens and the preparation of subunit vaccines. BALB / c mice were immunized with Toxoplasma gondii tachyzoite membrane protein and their spleen cells were fused with mouse myeloma cells SP2 / 0 to screen out hybridomas capable of stably secreting high titer anti-P30 monoclonal antibodies Cell lines, and the monoclonal antibody immunoglobulin subclasses and antibody titers were determined, the antigens recognized by the mAbs were localized with IFAT and identified by SDS-PAGE and Western Blot analysis. Results Two hybridoma cell lines, E3 and G2, against P30 antigen were obtained. The secreted antibodies did not cross-react with antigens such as Pneumocystis carinii and Cryptosporidium parvum. The two monoclonal antibodies belong to the Ig G1 subclass, and the recognized antigen is located on the tachyzoite surface membrane. The results show that the prepared anti-P30 antigen hybridoma cell lines can secrete high titers and specific monoclonal antibodies.