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    • 5. 发明授权
    • Method for producing influenza virus and vaccine
    • 生产流感病毒和疫苗的方法
    • US5698433A
    • 1997-12-16
    • US684729
    • 1996-07-22
    • Otfried KistnerNoel BarrettWolfgang MundtFriedrich Dorner
    • Otfried KistnerNoel BarrettWolfgang MundtFriedrich Dorner
    • A61K39/00A61K39/145A61K39/15A61K39/245C07K14/11C12N7/00C12N7/02
    • C07K14/005A61K39/12A61K39/145A61K39/15A61K39/245C12N7/00A61K2039/5252A61K2039/55505A61K39/00C12N2760/16122C12N2760/16134C12N2760/16151C12N2760/16234
    • A method of producing an influenza virus and vaccines derived from the virus utilizes cultured vertebrate biomass aggregates comprising a plurality of cell types derived from a plurality of vertebrate tissues and is particularly suitable for use with chicken embryo cultures. The method both eliminates the necessity to use costly methods requiring whole chicken embryos and provides proteases suitable for the activation of a wide variety of viruses. After infecting the cells of the culture with an influenza virus, which is preferably modified to create a cleavage site in the hemagglutinin of the virus, a substance such as a protease is introduced that cleaves the hemagglutinin. The culture then is incubated under conditions that permit growth of the virus. The method provides also for the augmentation of virus activation in the culture by the continuous or batchwise removal of culture media, treatment of the media with substances such as proteases which increase cellular activation, attenuation of any undesired effects of the augmentation and return of the augmented media to the culture. The vaccines produced from the harvested virus therefore are free of egg proteins and are much more economical to produce. The methodology of the present invention allows the large scale continuous production of many viruses to a high titre.
    • 生产流感病毒的方法和源自病毒的疫苗利用培养的脊椎动物生物质聚集体,其包含多种衍生自多个脊椎动物组织的细胞类型,并且特别适用于鸡胚培养。 该方法既消除了使用需要全部鸡胚胎的昂贵方法的必要性,并提供了适用于激活各种病毒的蛋白酶。 在用流感病毒感染培养物的细胞后,其优选被修饰以在病毒的血凝素中产生切割位点,引入了裂解血凝素的诸如蛋白酶的物质。 然后将培养物在允许病毒生长的条件下孵育。 该方法还用于通过连续或分批地去除培养基来增加培养物中的病毒活化,用诸如增加细胞活化的蛋白酶的物质处理培养基,减少增加的任何不期望的影响和增加的 媒体对文化。 因此,从收获的病毒产生的疫苗不含蛋蛋白,并且生产更经济。 本发明的方法允许大规模连续生产许多病毒到高滴度。