抗弓形虫表膜抗原 P30单克隆抗体的制备与鉴定(英文)

PREPARATION AND IDENTIFICATION OF MONOCLONAL ANTIBODIES AGAINST TOXOPLASMA MAJOR SURFACE ANTIGEN P30

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。

To develop and identify the monoclonal antibodies(McAbs) against the major surface antigen P30 of Toxoplasma gondii , which would be useful for further study. BALB/c mice were immunized with the surface antigen of tachyzoites. Spleen cells isolated from the immunized mice were fused with myeloma cell line SP2/0. After cell fusion, positive hybridoma cell lines were screened by ELISA. McAbs of E3 and G2 against the major surface antigen P30 were identified with SDS PAGE and Western Blot, and they were proved to be IgG1 subtypes by agar double diffusion technique. It was identified with IFAT that the antigens recognized by the McAbs were located at the surface of tachyzoite. The McAbs had no cross reactions with the antigens of Cryptosporidium、 Pneumocystis caranii and Cysticercus cellucose . The hybridoma cell lines secreting high McAb titers of antiP30 of T. gondii have been established.