摘要
目的 建立优化的人用H5N1禽流感病毒疫苗生产的工艺。方法 在不同的稀释倍数、收获时间及灭活剂添加量下,通过测量收获液的病毒滴度和血凝效价,来确定病毒的最佳生产条件,并对离心法和凝胶过滤层析法的纯化效果进行对比。结果 103~104半数鸡胚感染量(50% egg infective dose,EID50)病毒接种鸡胚,收获的鸡胚尿囊液的病毒滴度和血凝效价最高,分别为10-8.3EID50和1∶480;在56~72 h血凝效价最高。甲醛浓度1∶10 000灭活144 h为灭活最佳条件。两种纯化方法得到的样品纯度和卵清蛋白的去除率相近,但离心纯化法和凝胶过滤层析纯化法病毒回收率有较大的差异,分别为19%和70%。结论 成功建立了高产毒的鸡胚基质H5N1禽流感病毒培养、灭活及纯化工艺。
Objective To establish the optimized production process of human H5N1 avian influenza virus vaccine.Methods The optimal virus production conditions were determined by measuring virus titer and hemagglutination titer(HAT)under different dilution ratios,harvest time and amounts of inactivating agent.The purification effects of virus by centrifugation and gel filtration chromatography were also compared.Results When inoculated with 103-10450%egg infective dose(EID50)H5N1 avian influenza virus,the virus titer(10-8.3EID50)and HAT(1∶480)of allantoic fluid were the highest.The HATs were the highest at 56-72 h.The best inactivation condition was 1∶10000 formaldehyde for 144 h.The sample purity and the removal rate of ovalbumin were both similar between centrifugation and gel filtration chromatography purification methods,but the recovery rates of virus were 19%and 70%,respectively.Conclusion The culture,inactivation and purification process of H5N1 avian influenza virus in chicken embryo with high virus yield is successfully established.
作者
夏志武
施金荣
赵巍
孟子延
李兴航
刘博
季雅琪
杨晓明
Xia Zhiwu;Shi Jinrong;Zhao Wei;Meng Ziyan;Li Xinghang;Liu Bo;Ji Yaqi;Yang Xiaoming(National Engineering Technology Research Center of Combination Vaccines,Wuhan 430207,China;Department of Quality Control,Wuhan Institute of Biological Products Co.,Ltd.,Wuhan 430207,China;China National Biotec Group Co.,Ltd.,Beijing 100029,China)
出处
《国际生物制品学杂志》
CAS
2020年第2期62-65,共4页
International Journal of Biologicals
作者简介
通信作者:杨晓明,Email:yangxiaoming@sinopharm.com。
