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目的制备抗乙型流感单克隆抗体,为建立特异、灵敏、简便的乙型流感病毒快速检测方法奠定基础。方法应用鸡胚接种法、ELISA方法、免疫扩散法、血凝抑制法筛选能稳定分泌抗乙型流感病毒单抗的杂交瘤细胞,并对分泌的单抗的安全性,有效性、抗体亚型,特异性及灵敏性进行了全面评价。结果获得了3株能稳定分泌抗乙型流感病毒单抗的杂交瘤细胞株,分别命名为3G11、4F9、8G6;制备的抗乙型流感病毒单克隆抗体无鼠源病毒污染;抗体亚型为IgG2a亚型,轻链均为κ链;对乙型流感病毒具有高度特异性,而与甲型流感病毒、呼吸道合胞病毒、副流感病毒、呼吸道腺病毒等无交叉反应。结论制备的乙型流感病毒单克隆抗体具有特异性强、灵敏性高,为临床乙型流感的早期快速诊断及流行病调查奠定了实验基础。
Objective To prepare anti-influenza B monoclonal antibodies and lay a foundation for the establishment of a specific, sensitive and simple method for the rapid detection of influenza B virus. Methods The hybridoma cells secreting anti-influenza B virus monoclonal antibody were screened by the method of chicken embryo inoculation, ELISA, immunodiffusion and hemagglutination inhibition. The safety, efficacy, antibody subtype , Specificity and sensitivity of a comprehensive evaluation. Results Three hybridoma cell lines that can stably secrete anti-influenza B virus monoclonal antibodies were obtained and named as 3G11, 4F9 and 8G6, respectively. The monoclonal antibodies against influenza B virus were not contaminated by mouse viruses. The antibody subtypes were IgG2a subtype and light chain are both κ chain. It has high specificity for influenza B virus but no cross-reaction with influenza A virus, respiratory syncytial virus, parainfluenza virus and respiratory adenovirus. Conclusion The prepared monoclonal antibody of influenza B virus has strong specificity and high sensitivity, which lays the foundation for early rapid diagnosis and epidemiological investigation of clinical influenza B.