摘要
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。
To develop a specific,sensitive and rapid method for detection of influenza B virus,we prepared and evaluated a monoclonal antibodies（mAbs） against influenza B virus in this study.By using egg inoculation method,ELISA,double immunodiffusion,and hemagglutination inhibition assay,we screened hybridoma cell lines and found out three hybridoma cell lines which could steadily secret anti-influenza B virus monoclonal antibody,named as 3G11,4F9,and 8G6 respectively.Meanwhile the safety,efficacy,antibody subtype,specificity,and sensitivity of these monoclonal antibodies were evaluated.These results showed that the mAbs were characterized as IgG2a subclasses,of which all the light chains were κ chains.Among them no murine viruses exist.More important,these monoclonal antibodies could specifically detected all of the influenza B viruses tested,but did not react with the influenza H1N1 virus,influenza H3N2 virus,respiratory syncytial virus,parainfluenza virus type I/III,or adenovirus 3/7 virus.These findings demonstrated that the influenza B virus monoclonal antibodies will provide sensitive and robust tools for rapid diagnosis,on-site detection,and epidemiological investigation of clinical influenza B virus infection.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2012年第12期1077-1080,共4页
Immunological Journal
