刚地弓形虫表面蛋白SAG1单克隆抗体的制备及鉴定
首发时间:2016-01-18
摘要:用刚地弓形虫表面SAG1免疫BALB/c小鼠后,将其脾细胞与骨髓瘤细胞SP2/0进行融合,获得7株抗刚地弓形虫SAG1单克隆抗体4H3、4F9F6、7D8D7、2A12G10、2A12H10、8D8E8和1G10A7。4H3、2A12G10和2A12H10亚型为IgG1,4F9F6、7D8D7、8D8E8和1G10A7亚型为IgG2a。4F9F6、7D8D7和8D8E8可以特异性与弓形虫速殖子表面识别。4F9F6抗体与SAG1抗原亲和力高于7D8D7抗体和8D8E8抗体,其亲和常数Kd为6.09×10-10。7D8D7和8D8E8抗体的亲和常数Kd分别为7.03×10-10和2.47×10-9。本研究成功制备了3株具有高亲和力的抗刚地弓形虫单克隆抗体,为抗刚地弓形虫抗体的人源化抗体奠定基础。
关键词: 关键词:刚地弓形虫 速殖子表面蛋白SAG1 单克隆抗体 生物学特性
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Preparation and identification of monoclonal antibodies angaist the surface antigen SAG1 of Toxoplasma gondii
Abstract:Abstract: After the BALB/c mice was immunized by the surface antigen SAG1 of Toxoplasma gondii, the spleen cells of mice was fused with with the myeloma cells (SP2/0). Seven hybridoma cell lines secreting monoclonal antibody (McAb) against the surface antigen SAG1 were obtained, and named as 4H3, 4F9F6, 7D8D7, 2A12G10, 2A12H10, 8D8E8 and 1G10A7. The isotype of 4H3, 2A12G10 and 2A12H10 belonged to IgG1, and the isotype of 4F9F6, 7D8D7, 8D8E8 and 1G10A7 belonged to IgG2a. The antibody of 4F9F6, 7D8D7 and 8D8E8 can recongize specifically with the tachyzoite of T. gondii. The affinity of 4F9F6 antibody with SAG1 was higher than that of 7D8D7 antibody and 8D8E8 antibody, and the affinity constant Kd was 6.09×10-10. The three McAbs angaist the SAG1 of T. gondii in this study can be used to produce the humanzied antibody agaist Toxoplasmosis.
Keywords: Key words: Toxoplasma gondii the suface antigen 1 (SAG1) of the tachyzoite monoclonal antibody the biological characteristics
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No.4675845568507145****
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