猪胸膜肺炎放线杆菌单克隆抗体的制备与鉴定

Preparation and Identification of the Monoclonal Antibodies of Actinobacillus Pleuropneumoniae in Swine

目的:建立抗血清1型猪胸膜肺炎放线杆菌(APP-1)脂多糖(LPS)的单克隆抗体杂交瘤细胞株,为今后建立该菌的免疫检测技术奠定基础。方法:腹腔注射APP-1标准株的甲醛固定抗原免疫BALB/c小鼠,ELISA检测血清中LPS抗体滴度并选择值最高的小鼠用于细胞融合,且于融合前3天加强免疫1次。常规方法进行融合,HT、HAT培养液选择培养融合后的细胞,ELISA检测细胞培养上清中的LPS抗体,阳性杂交瘤细胞用有限稀释法克隆传代。结果:获得2株稳定分泌抗APP-1LPS抗原的单克隆抗体杂交瘤细胞株,其小鼠腹水单克隆抗体效价132000～164000。结论:已成功建立2株稳定分泌抗APP-1LPS抗原的单克隆抗体杂交瘤细胞株,为今后建立该菌的免疫胶体金分型诊断技术奠定基础。

Objective： To establish actinobacillus pleuropneumoniae Type 1 antiserum Actinobacillus actinomycetemcomitans （APP-1） lipopolysaccharide （LPS） monoclonal antibody hybridoma cell lines and to lay a foundation for immunity examination technology for this bacteria. Methods： Intraperitoneal injection of formalin-fixed APP-1 strain vaccine antigen,ELISA antibody titer of serum LPS and choose the mouse with the highest value for cell fusion. Conduct an enhancing immunization three days before Conventional methods of integration. Selectively cultivate fusion cells in HT and HAT medium, Cell culture supematant is checked for LPS antibody using ELISA. Positive hybridoma cells are cloned using limited dilution. Results： 2 swains secreting anti APP- 1 antigen monoclonal antibody hybridoma cell lines of LPS are attained. Mouse ascites monoclonal antibody titer is 1： 32 000 - 1：64 000. Conclusion： Three swains secreting anti - LPS APP- 1 antigen monoclonal antibody hybridoma cell lines are successfully established, which lay a foundation for future differential diagnosis of colloidal gold technologies for this bacteria. The bacteria build for the future differential diagnosis of colloidal gold technologies.