抗胸膜肺炎放线杆菌毒素Ⅰ、Ⅱ和Ⅲ的单克隆抗体的制备  

Generation and Characterization of the Monoclonal Antibodies Against ApxⅠ,ApxⅡ and ApxⅢ of Actionbacillus pleuroneumoniae

作　　者：黄红亮[1] 李冲[1] 周锐[1] 谢甜甜[1] 陈美玲[1] 魏洁[1] 洪文洲[1] 陈焕春[1] 

机构地区：[1]华中农业大学动物医学学院,武汉430070

出　　处：《农业生物技术学报》2005年第5期659-663,共5页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(No.30200011);中国博士后科学基金(No.20040350179)资助

摘　　要：分别用大肠杆菌(Escherichia coli)表达的猪传染性胸膜肺炎放线杆菌(A ctinobacillus pleuropneum oniae)外毒素(A px)Ⅰ、Ⅱ和Ⅲ为抗原,免疫Balb/c小鼠,经细胞融合、克隆与筛选,分别获得能稳定分泌A pxⅠ、A pxⅡ和A pxⅢ单克隆抗体的杂交瘤2株(4、16)、3株(14、3F11、16G10)和2株(44、53)。这7株杂交瘤细胞的染色体数分别为87(81￣94)、84(81￣88)、90(85￣94)、87(80￣94)、82(79￣85)、93(88￣99)和85(82￣87),其腹水ELISA效价分别为100×210、100×29、100×210、100×27、100×214、100×214和100×212。间接ELISA和W estern blot分析结果证明,所有单克隆抗体均具有与相应天然毒素的反应活性,且特异性良好。Seven hybridoma clones were generated by fusion of the SP2/0 myeloma cells and the splcnocytcs obtained from Balb/c mice immunized by the recombinant Apx Ⅰ, Apx Ⅱ and Apx Ⅲ of Actionbacillus pleuropneumoniae （APP）. Among the 7 hybridoma clones, 2 of them （4 and 16） could secret monoclonal antibodies against ApxI, 3 of them （3F11,16G10 and 14） against Apx Ⅱ and the rest 2 （44 and 53） against Apx Ⅲ. Their chromosome numbers were 81～94, 81～88, 85～94, 80～94, 79～85, 88～99 and 82～87, respectively. The ELISA titers of the ascite fluids induced by the 7 hybridomas were 100×2^10, 100×2^9, 100×2^10, 100×2^7, 100×2^14, 100×2^14 and 100×2^12, respectively. Indirect ELISA and Western blotting analyses indicated that all the 7 strains of monoclonal antibodies could react specifically with the corresponding Apx toxin isolated from the cultures of different APP strains.

关 键 词：胸膜肺炎放线杆菌 ApxⅠ ApxⅡ ApxⅢ 单克隆抗体 

分 类 号：S858.28[农业科学—临床兽医学] S852.43[农业科学—兽医学]