流感病毒经典重配中X-157母本株HA抗血清制备

摘要

Objective Preparation of maternal strain X-157 HA antiserum for influenza virus classical reassortment.Methods X-157 virus was digested by bromelain after inactivation and purification.Sucrose continuous density gradient centrifugation method was used to purify HA protein.SDS-PAGE method was used to select the right protein band.Rabbit antiserum was collected after immunization.Antiserum HI titer was identified by HI method.Antiserum purity was identified by SIRD method.Results High Immunogenic X-157 HA protein was successfully prepared and the titer of anti-HA protein by HI reached to 2560.High purity HA antiserum was identified by SIRD method.Conclusions 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.%目的 制备流感病毒经典重配过程中所需的X-157母本株血凝素蛋白(Hemagglutinin,HA)抗血清.方法 X-157株流感病毒灭活纯化后经菠萝蛋白酶消化HA蛋白,蔗糖连续密度梯度离心法纯化,SDS-PAGE法鉴定纯化HA蛋白,免疫兔子收集抗血清,血凝抑制试验法(Hemoglutination Inhibition assay,HI)测定抗血清效价,单向免疫扩散法(Single Radial Immunodiffusion,SIRD)鉴定HA抗血清纯度.结果 成功制备了高免疫原性X-157株HA蛋白,兔抗HA血清HI滴度可达2560,SIRD鉴定为高纯度HA抗血清.结论 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.