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发明公开
CN108570445A PK15细胞驯化悬浮方法和二阶病毒生产工艺 审中-实审 转让

PK15细胞驯化悬浮方法和二阶病毒生产工艺
摘要：
本发明涉及PK15细胞驯化悬浮方法和二阶病毒生产工艺。具体而言，本发明涉及驯化PK15细胞使其适应无血清悬浮培养，并以其为宿主细胞，采用二阶培养接毒方式生产能在悬浮PK15细胞上病变或有敏感性的病毒，即，细胞长至2.0～20.0×106细胞/mL时，用生产培养基稀释1～5倍，使密度在1.0～10.0×106细胞/mL间，然后接毒，或在稀释前接毒。本发明的悬浮PK15细胞适应在无血清和化学成分确定的培养基中悬浮生长，其二阶病毒生产工艺简单，易放大，不换液，培养基利用率高；生产和生长培养基可相同或不同，二者按照比例混合，即可使细胞二次生长并促进病毒表达。
摘要（英）：

The invention relates to a PK15 cell acclimation suspension method and a two-stage virus production process. According to the PK15 cell acclimation suspension method, PK15 cells are acclimated to adapt to serum-free suspension culture and used as host cells for producing viruses which can have cytopathic effect or sensitivity on suspension PK15 cells by two-stage culture virus inoculation; when the cells grow to 2.0-20.0*10<6> cells/mL, diluting with a production culture medium by 1-5 times is performed to reach a density range of 1.0-10.0*10<6> cells/mL, and then virus inoculation is carriedout or virus inoculation is performed before dilution. The suspension PK15 cells are adaptive to suspension growth in the serum-free chemical-defined culture medium, and the two-stage virus productionprocess is simple and easy in amplification; liquid changing is avoided, and high culture medium utilization efficiency is achieved; production and growth culture media can be identical or not and can be mixed proportionally to realize cell secondary growth and viral expression promotion.

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