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    • 6. 发明申请
    • VIRUS PURIFICATION METHODS
    • 病毒净化方法
    • WO2005080556A9
    • 2006-10-26
    • PCT/EP2005050739
    • 2005-02-21
    • CRUCELL HOLLAND BVWEGGEMAN MIRANDAVAN CORVEN EMILE JOANNES JOSEP
    • WEGGEMAN MIRANDAVAN CORVEN EMILE JOANNES JOSEP
    • C12N7/02
    • C12N7/00C12N2501/70C12N2710/10051
    • The invention provides a method for the purification of a virus from a host cell, said method comprising the steps of: a) culturing host cells, b) infecting said host cells with a virus, c) treating the cell culture with nuclease, d) lysing said host cells to provide a lysate comprising the virus. The virus is preferably a recombinant adenovirus. The invention further provides a method for the purification of a recombinant virus expressing a heterologous protein that is capable of binding nucleic acid, comprising the steps of: a) culturing host cells, b) infecting said host cells with said recombinant virus, c) lysing said host cells to provide a lysate comprising said recombinant virus, d) subjecting the recombinant virus to anion exchange chromatography and size exclusion chromatography, characterized in that the virus containing mixture is buffer exchanged at least once with a solution comprising at least 2 M NaCl, or another salt providing an equivalent ionic strength.
    • 本发明提供了从宿主细胞纯化病毒的方法,所述方法包括以下步骤:a)培养宿主细胞,b)用病毒感染所述宿主细胞,c)用核酸酶处理细胞培养物,d) 裂解所述宿主细胞以提供包含病毒的裂解物。 该病毒优选是重组腺病毒。 本发明进一步提供了纯化表达能够结合核酸的异源蛋白质的重组病毒的方法,包括以下步骤:a)培养宿主细胞,b)用所述重组病毒感染所述宿主细胞,c)裂解 所述宿主细胞提供包含所述重组病毒的裂解物,d)使所述重组病毒经历阴离子交换层析和尺寸排阻层析,其特征在于,所述含病毒混合物用包含至少2M NaCl的溶液缓冲液交换至少一次, 或提供相等离子强度的另一种盐。