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    Mammalian ceII culture processes for protein production
    4.
    发明申请
    Mammalian ceII culture processes for protein production 有权
    用于蛋白质生产的哺乳动物ceII培养方法

    公开(公告)号：US20050019859A1

    公开(公告)日：2005-01-27

    申请号：US10742564

    申请日：2003-12-18

    申请人： Bernhard Schilling ,  Linda Matlock ,  Stephen Zegarelli ,  William Burnett ,  Christoph Joosten ,  Jonathan Basch ,  Sivakesava Sakhamuri ,  Steven Lee

    发明人： Bernhard Schilling ,  Linda Matlock ,  Stephen Zegarelli ,  William Burnett ,  Christoph Joosten ,  Jonathan Basch ,  Sivakesava Sakhamuri ,  Steven Lee

    IPC分类号： C07K14/705 ,  C12N5/00 ,  C12N5/02 ,  C12P21/00 ,  C07K14/74

    CPC分类号： C12P21/005 ,  C07K14/70521 ,  C12N5/0018 ,  C12N2500/34 ,  C12N2500/74 ,  C12N2501/33 ,  C12N2501/90 ,  C12N2501/905 ,  C12N2501/91 ,  C12N2510/02

    摘要： The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, preferably, but not limited to, fed-batch cell cultures. In one aspect, the methods comprise at least two temperature shifts performed during the culturing period, in which the temperature is lower at the end of the culturing period than at the time of initial cell culture. Throughout their duration, the culturing processes of the invention involving two or more downward shifts in temperature sustain a high viability of the cultured cells, and can yield an increased end titer of protein product, and a high quality of protein product, as determined, e.g., by sialic acid content of the produced protein. In another aspect, the methods comprise the delayed addition of polyanionic compound during the culturing period. The delayed addition of polyanionic compound sustains a high viability of the cultured cells, and can extend the growth phase, delay the onset of the death phase, and arrest the death phase.

    摘要翻译： 本发明描述了通过动物细胞或哺乳动物细胞培养物生产蛋白质，特别是糖蛋白的方法和方法，优选但不限于补料分批细胞培养物。 在一个方面，所述方法包括在培养期间进行的至少两个温度漂移，其中在培养期结束时温度比初始细胞培养时的温度更低。 在整个持续时间内，涉及两个或更多个温度向下移位的本发明的培养方法维持培养细胞的高活力，并且可以产生蛋白质产物的最终滴定度和蛋白质产物的高质量，如所确定的，例如 ，通过产生蛋白质的唾液酸含量。 另一方面，所述方法包括在培养期间延迟加入聚阴离子化合物。 聚阴离子化合物的延迟添加维持培养细胞的高活力，并且可以延长生长期，延缓死亡阶段的发作并停止死亡期。

    Product quality enhancement in mammalian cell culture processes for protein production
    6.
    发明申请
    Product quality enhancement in mammalian cell culture processes for protein production 有权
    用于蛋白质生产的哺乳动物细胞培养过程中的产品质量增强

    公开(公告)号：US20050084933A1

    公开(公告)日：2005-04-21

    申请号：US10740645

    申请日：2003-12-18

    申请人： Bernhard Schilling ,  Scott Gangloff ,  Dharti Kothari ,  Kirk Leister ,  Linda Matlock ,  Stephen Zegarelli ,  Christoph Joosten ,  Jonathan Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

    发明人： Bernhard Schilling ,  Scott Gangloff ,  Dharti Kothari ,  Kirk Leister ,  Linda Matlock ,  Stephen Zegarelli ,  Christoph Joosten ,  Jonathan Basch ,  Sivakesava Sakhamuri ,  Steven S. Lee

    IPC分类号： C07K14/705 ,  C12P21/00 ,  C12P21/02 ,  C12N5/06

    CPC分类号： C07K14/70521 ,  C12N2500/34 ,  C12P21/005

    摘要： The present invention describes methods and processes for the production of proteins, particularly glycoproteins, by animal cell or mammalian cell culture, illustratively, but not limited to, fed-batch cell cultures. The methods comprise feeding the cells with D-galactose, preferably with feed medium containing D-galactose, preferably daily, to sustain a sialylation effective level of D-galactose in the culture for its duration, thus increasing sialylation of the produced proteins. The methods can also comprise at least two temperature shifts performed during the culturing period, in which the temperature is lower at the end of the culturing period than at the time of initial cell culture. The cell culture processes of the invention involving two or more temperature shifts sustain a high cell viability, and can allow for an extended protein production phase. The methods can also comprise the delayed addition of polyanionic compound at a time after innoculation. Supplementation of the cultures with D-galactose, preferably in a feed medium, to sustain galactose at sialylation effective levels in the cultures until the end of a culture run reverses a decline in sialylation that accompanies culture scale up, and is advantageous for large scale culturing processes.

    摘要翻译： 本发明描述了通过动物细胞或哺乳动物细胞培养物生产蛋白质，特别是糖蛋白的方法和方法，例如但不限于补料分批细胞培养物。 所述方法包括用D-半乳糖喂养细胞，优选用含有D-半乳糖的饲料培养基，优选每天喂养，以维持培养基中唾液酸化有效水平的D-半乳糖的持续时间，从而增加产生的蛋白质的唾液酸化。 所述方法还可以包括在培养期间进行的至少两个温度漂移，其中在培养期结束时温度比初始细胞培养时温度更低。 涉及两个或多个温度变化的本发明的细胞培养方法维持高的细胞活力，并且可以允许延长的蛋白质生产阶段。 所述方法还可以包括在接种后延迟加入聚阴离子化合物。 用D-半乳糖，优选在饲料培养基中补充培养物，以维持培养物中唾液酸化有效水平的半乳糖，直至培养结束时反转伴随培养物扩大的唾液酸化的下降，并且有利于大规模培养 过程。