流感病毒A/PR/8/34重配母本株高免疫原性HA蛋白制备及鉴定  

Preparation and Identification of High Immunogenic A/PR/8/34 Maternal Strain HA Protein for Influenza Virus Classical Reassortment

作　　者：唐静[1] 辛丽[1] 郭俊峰[1] 朱闻斐[1] 张贺源 郎绍辉 王大燕[1] 舒跃龙[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所卫生部医学病毒和病毒病重点实验室,北京102206

出　　处：《病毒学报》2016年第2期141-144,共4页Chinese Journal of Virology

基　　金：重大专项"艾滋病和病毒性肝炎等重大传染病防治"-疫苗新型细胞基质;佐剂和免疫递送系统研究(2013ZX10004003);重大专项"重大新药创造"-流感疫苗应急研发体系能力建设及产品研发(2015ZX09101044)

摘　　要：制备流感病毒经典重配过程中所需的A/PR/8/34母本株血凝素蛋白（Hemagglutinin,HA）抗血清。A/PR/8/34流感病毒灭活纯化后经菠萝蛋白酶消化。5%~20%蔗糖连续密度梯度离心法纯化HA蛋白。单向免疫扩散法（Single Radial Immunodiffusion,SRID）筛选目的蛋白。SDS-PAGE法鉴定目的蛋白。纯化的A/PR/8/34 HA蛋白免疫兔子制备抗血清,血凝抑制试验（Hemoglutination Inhibition Assay,HI）测定HA抗血清效价,经SRID法鉴定HA抗血清纯度。成功制备了高免疫原性A/PR/8/34HA蛋白;兔抗血清HI滴度可达10240,SRID法鉴定为高纯度HA抗血清。制备了流感病毒经典重配中关键试剂,摸索成功整套技术方法,为我国自主研发季节性流感疫苗株奠定了基础。Preparation of maternal strain A/PR/8/34 HA antiserum for influenza virus classical reassortment.A/PR/8/34 virus was digested by bromelain after inactivation and purification.5%~20% sucrose continuous density gradient centrifugation method was used to purify HA protein.SIRD method was used to select the target protein.SDS-PAGE method was used to identified HA protein.High Immunogenic A/PR/8/34 HA protein was successfully prepared and HI titer reached 10240.High purity HA antiserum was identified by SIRD method.The key reagent in the classical reassortment of influenza virus was prepared,and the complete set of technical methods were explored,which laid the foundation for the independent research and development of seasonal influenza vaccine strains of China.

关 键 词：流感病毒 经典重配 HA蛋白 高免疫原性 HA抗血清 

分 类 号：R373.1[医药卫生—病原生物学]