畜牧与兽医2026，Vol.58Issue(9)：114-119,6.
抗副鸡禽杆菌单克隆抗体的制备与鉴定
Preparation and characterization of monoclonal antibodies against Avibacterium paragallinarum
摘要
Abstract
This study aimed to prepare monoclonal antibodies(mAbs)against Avibacterium paragallinarum(AP).Whole-cell inactivated AP bacteria were used as the immunogen to immunize 6-to 8-week-old BALB/c mice.After three immunizations,spleen cells from the im-munized mice were collected and fused with SP2/0 myeloma cells.An indirect ELISA method using whole AP bacteria as the coating antigen was established for screening.Positive hybridoma cells were subjected to three rounds of subcloning,and the obtained mAbs were character-ized by indirect ELISA,antibody subclass identification,Western blot,and indirect immunofluorescence assay.Three hybridoma cell lines that could stably secrete AP-specific mAbs were successfully obtained and designated as 1B4,1F9,and 3C11.The mAbs secreted by these cell lines were identified as IgG3,IgG3,and IgG1 subclasses,respectively.ELISA results showed that the antibody titers in hybridoma cul-ture supernatants were 1∶512 for clones 1B4 and 1F9,and 1∶256 for clone 3C11.The corresponding titers in ascites fluid were 1∶204 800 for clones 1B4 and 1F9,and 1∶102 400 for clone 3C11.Western blot analysis revealed that all three mAbs could recognize AP bacterial proteins:1B4 recognized a protein of approximately 39 kDa,while both 1F9 and 3C11 recognized a protein of approximately 36 kDa.Indirect immunofluorescence assay further confirmed that all three mAbs could react specifically with the three AP strains tested,producing character-istic fluorescence.In conclusion,the antiAP monoclonal antibodies were successfully prepared in this study,and the target proteins they rec-ognize were preliminarily identified,providing important biological materials for the future establishment of detection methods and studies on the pathogenic mechanisms of AP.关键词
副鸡禽杆菌/单克隆抗体/全菌免疫Key words
Avibacterium paragallinarum/monoclonal antibody/whole-cell immunization分类
农业科技引用本文复制引用
潘新语,丁香,宋超逸,卞鸿凯,陈佳和,王海燕,刘文博..抗副鸡禽杆菌单克隆抗体的制备与鉴定[J].畜牧与兽医,2026,58(9):114-119,6.基金项目
江苏省优势学科第四期项目(2021) （2021）
