肠道病毒71型鼠源性中和单克隆抗体的制备  

Preparation of murine monoclonal antibody with neutralizing effect on enterovirus 71

作　　者：朱文兵[1] 严丽蔚 巩蔚[1] 张雪梅[1] 吴忠香[1] 徐婧雯[1] 宋杰[1] 董少忠[1] ZHU Wen-bing, YAN Li-wei, GONG Wei, ZHANG Xue-mei, WU Zhong-xiang, XU Jing-wen, SONG Jie, DONG Shao-zhong(Institute of Medical Biology, Chinese Academy of Medical Science and Peking Union Medical College, Kunming 650118, Yunnan Province, Chin)

机构地区：[1]中国医学科学院北京协和医学院医学生物学研究所云南省重大传染病疫苗研发重点实验室,云南昆明650118

出　　处：《中国生物制品学杂志》2018年第3期257-261,共5页Chinese Journal of Biologicals

基　　金：云南省科技厅重点新产品研发计划(2015BC007)

摘　　要：目的制备肠道病毒71型(enterovirus A 71,EV71)鼠源性中和单克隆抗体。方法以灭活EV71病毒液为免疫原免疫BALB/c小鼠,通过杂交瘤技术获得抗EV71单克隆抗体;ELISA法检测抗体的特征,获得具有高效结合性的单克隆抗体;SDS-PAGE和Western blot法鉴定Protein A纯化的抗体;通过体外和体内中和试验鉴定抗体的中和特性。结果得到3株针对EV71 VP1的高效结合性单克隆抗体15H7、15G7和6G2,且15H7经中和试验证明具有较强的中和活性。结论成功制备了EV71鼠源性中和单克隆抗体,其在小鼠体内能成功地抑制病毒增殖。Objective To prepare the murine monoclonal antibody（McAb） with neutralizing effect on enterovirus 71（EV71）. Methods BALB/c mice were immunized with inactivated EV71 to prepare McAb by hybridoma technique.The McAb with high affinity was screened by ELISA. The McAb purified with protein A was identified by SDS-PAGE and Western blot. The neutralizing property of McAb was identified by in vitro and in vivo neutralization tests. Results Monoclonal antibodies with high affinity,15H7,15G7 and 6G2,were prepared,of which 15H7 showed high neutralizing activity as proved by neutralization test. Conclusion Murine McAb against EV71 was prepared,which inhibited the virus proliferation in mice successfully.

关 键 词：肠道病毒71型 单克隆抗体 中和作用 

分 类 号：R373.2[医药卫生—病原生物学] R392-33[医药卫生—基础医学]