摘要
目的建立四价流感病毒亚单位疫苗制备方法。方法将4株毒种分别接种于9~11日龄的健康鸡胚尿囊腔中,在33~35℃培养箱培养48~72 h后,冷胚处理,收获尿囊液,经澄清、病毒灭活、裂解和纯化,除菌过滤制成单价疫苗原液;利用聚丙烯酰胺凝胶电泳(SDS-PAGE)方法,分析有效成分及纯度。将收获的4种单价疫苗原液进行混配,制成四价流感病毒亚单位疫苗。最终按《中华人民共和国药典》2015年版对成品疫苗进行全面检定。结果成功制备出四价流感病毒亚单位疫苗,其有效成分血凝素含量占总蛋白比例为91.4%。4种疫苗株血凝素含量分别为H1N1:30μg/ml、H3N2:34μg/ml、B1:36μg/ml、B2:30μg/ml,对成品的检定完全合格。结论本研究所制备的四价流感亚单位疫苗符合生产和检定规程要求。
Objective To prepare the 4-valent influenza virus subunit vaccine.Methods Four strains of virus were inoculated into the allantoic cavity of healthy chicken embryo which were 9 to 11 days old.After incubated at 33-35℃for 48-72 h,the embryos were cooled and the allantoic fluid was harvested.Following virus inactivation,lysis and purification,sterilization and filtration were applied to prepare a monovalent vaccine stock solution;polyacrylamide gel electrophoresis(SDS-PAGE)was used to analyze the purity.The harvested four monovalent stocks are mixed to prepare a tetravalent influenza virus subunit vaccine.Finally,the finished product vaccine was fully verified according to the Pharmacopoeia of the People’s Republic of China(2015 edition).Results The tetravalent influenza virus subunit vaccine was successfully prepared,among which hemagglutinin accounted for 91.4%of the total protein.The hemagglutinin of the four vaccine strains were H1N1 for 30μg/ml,H3N2 for 34μg/ml,B1 for 36μg/ml,and B2 for 30μg/ml,respectively.Conclusion The four-valent influenza subunit vaccine produced meets the requirements of production and verification.
作者
李雪峰
陈宁
徐朝静
姚蕊
阮承迈
Chen Ning;Li Xuefeng;Xu Zhaojing;Yao Rui;Ruan Chengmai(Department Research and Development,Zhongyianke Biotechnology Co.,Ltd.Tianjin 300300,China)
出处
《中华实验和临床感染病杂志（电子版）》
CAS
2019年第6期524-527,共4页
Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)
基金
天津市科技计划项目(No.17ZXSCSY00010)
关键词
四价流感病毒亚单位疫苗
四价
流感
疫苗
乙型
Tetravalent influenza virus subunit vaccine
Influenza
Vaccine
Type B influenza virus
