抗迟缓爱德华氏菌单克隆抗体制备及初步鉴定

Preparation and characterization of monoclonal antibody against Edwardsiella tarda

用灭活迟缓爱德华氏菌菌株AL60306NA1免疫Balb/c小鼠,采用杂交瘤技术制备抗迟缓爱德华氏菌的McAb,获得3株可持续分泌抗迟缓爱德华氏菌McAb的杂交瘤细胞株3A7、4F11和4C9;腹水效价分别达到1.0×10-6、1.0×10-6和1.0×10-5;细胞亚型分别为IgG1、IgG2b、IgM.交叉反应结果显示,3A7、4F11、4C9与常见水产病原菌均无交叉反应;对菌株AL60306NA1检测灵敏度分别为1.25×104、1.25×105和1.25×105cfu.mL-1.结果表明,建立的3株杂交瘤细胞株所分泌抗迟缓爱德华氏菌McAb的效价高、特异性好、检测敏感性强,不仅可应用于迟缓爱德华氏菌的快速检测,也为迟缓爱德华氏菌免疫保护研究和疫苗制备奠定了基础.

The Edwardsiella tarda strain AL60306NA1 isolated from European eel was inactivated by formaldehyde and used as antigen to immunizate Balb/c mice.Three hybridomas strains secreting monoclonal antibodies（McAb） against Edwardsiella tarda were obtained and named 3A7、 4F11、 4C9.The titers of these 3 McAbs in ascites of Balb/c mice were 1.0×10-6、 1.0×10-6、 1.0×10-5,and the isotype were IgG1、 IgG2b、 IgM respectively.The sensitivity of McAbs were 1.25×104、 1.25×105、 1.25×105 cfu·mL-1 by using inacticated E.tarda cells as antigen.The McAb had no cross reactions with 7 other aquatic pathogenic bacteria.The specificity and sensitivity of E.tarda McAbs could be used for the rapid detction of edwardsiellosis in the future.