摘要
传染性法氏囊病毒(IBDV)VP2蛋白具有良好的免疫源性。本研究用纯化的IBDV NN1172株与弗氏佐剂混合,经腹腔注射途径免疫5周龄BALB/c小鼠,3次免疫后,对小鼠进行眼眶采血,用ELISA方法鉴定VP2抗体水平,用抗体水平最高小鼠的脾细胞与SP2/0进行融合后,对融合成功的杂交瘤细胞经亚克隆后获得1株传代稳定的单克隆细胞株-2G5(重链IgG2a型,轻链为κ型),经ELISA、IFA及Western blot验证结果表明,该细胞株产生的IBDV VP2蛋白抗体效价高、特异性强。本研究为IBDV VP2蛋白表达检测提供了生物材料。
The VP2 protein is the main structural protein of infectious bursal disease virus(IBDV)that shows good immunogenicity.In this study,five-week-old BALB/c mice were immunized three times with the purified IBDV strain NN1172 mixed with the adjuvant.The antibody level of VP2 was evaluated by ELISA.The spleen cells of the mice with the highest antibody level were selected for fusion and the hybridomas were screened by ELISA,IFA and Western blot.The monoclonal antibody 2G5 specific to VP2 was obtained through 3 cycles of limited dilution subcloning.The mAb 2G5 belonged to heavy chain IgG2a and light chainκ.The availability of a specific mAb to VP2 provided a useful tool for detection of IBDV.
作者
段绪来
刘静宜
郭伟强
陈金南
王致远
何秀苗
陈鸿军
DUAN Xulai;LIU Jingyi;GUO Weiqiang;CHEN Jinnan;WANG Zhiyuan;HE Xiumiao;CHEN Hongjun(Guangxi Key Laboratory for Polysaccharide Materials and Modifications,School of Marine Sciences and Biotechnology,Guangxi University for Nationalities,Nanning 530008,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)
出处
《中国动物传染病学报》
北大核心
2025年第4期184-189,共6页
Chinese Journal of Animal Infectious Diseases
基金
国家自然科学基金项目(32160824)
十四五国家重点研发专项(2022YFD1801000)。
作者简介
第一作者:段绪来,男,硕士研究生,生物化学与分子生物学专业;通信作者:何秀苗,E-mail:hxm@gxmzu.edu.cn;通信作者:陈鸿军,E-mail:vetchj@shvri.ac.cn。
