抗破伤风类毒素单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibodies against tetanus toxoid

作　　者：董国霞[1] 张华捷[1] 晁哲[1] 田霖[1] 侯启明[1] 谭亚军[1] 马霄[1] DONG Guo-Xia;ZHANG Hua-Jie;CHAO Zhe;TIAN Lin;HOU Qi-Ming;TAN Ya-Jun;MA Xiao(Division of DTP Vaccine and Toxins,Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products,National Institute for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院百白破疫苗和毒素室,卫生部生物技术产品检定方法及其标准化重点实验室,北京102629

出　　处：《中国免疫学杂志》2021年第18期2252-2256,共5页Chinese Journal of Immunology

基　　金：“十三五”国家科技重大专项(2018ZX09738005)项目。

摘　　要：目的:制备抗破伤风类毒素单克隆抗体,并对其进行鉴定。方法:用破伤风类毒素免疫BALB/c小鼠,经细胞融合及筛选,将阳性杂交瘤细胞株扩大培养后免疫BALB/c小鼠,收获的腹水用Protein G柱纯化;对获得的单克隆抗体进行鉴定,对杂交瘤细胞株进行稳定性检测。结果:筛选出5株稳定分泌抗破伤风类毒素单克隆抗体的杂交瘤细胞株,分别命名为T1、T2、T3、T4、T5,其分泌的单克隆抗体分别为IgG1、IgG2b、IgG1、IgG2b和IgG1亚类,轻链均为к型;均与百白破疫苗中的T抗原具有较好的反应,与D抗原、PT抗原、FHA抗原和PRN抗原无交叉反应;其中2株单克隆抗体为中和性抗体。结论:获得5株抗破伤风类毒素单克隆抗体,其中2株为中和性抗体,为破伤风疫苗效价检测新方法的建立等奠定了基础。Objective:To prepare and identify monoclonal antibodies against tetanus toxoid.Methods:BALB/c mice were immunized with tetanus toxoid,cells were fused and the positive hybridoma cell strains secreting tetanus antibodies were screened out.After the positive hybridoma cell strains were cultured on expansion,BALB/c mice were immunized,ascites were harvested and purified by Protein G-column,and monoclonal antibodies were obtained.The monoclonal antibodies were identified and the hybridoma cell strains were tested for stability.Results:Five hybridoma cell strains secreting monoclonal antibodies against tetanus toxoid were screened out,named T1,T2,T3,T4 and T5 respectively.The monoclonal antibodies were IgG1,IgG2b,IgG1,IgG2b and IgG1 subclasses respectively and all light chains wereкtype.All of them had good reaction with T antigen and had no cross reaction with D antigen,PT antigen,FHA antigen and PRN antigen in DTP vaccine.Two of them were neutralizing antibodies.Conclusion:Five monoclonal antibodies against tetanus toxoid have been obtained and two of them are neutralizing antibodies,which provides the foundation for the new method establishment of the potency test of tetanus vaccine etc.

关 键 词：破伤风类毒素 单克隆抗体 制备 鉴定 

分 类 号：R392.1[医药卫生—免疫学]