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1. 发明申请

US20050084933A1 Product quality enhancement in mammalian cell culture processes for protein production 有权
标题翻译：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。

2. 发明申请

US20080070280A1 Product quality enhancement in mammalian cell culture processes for protein production 审中-公开
公开(公告)号：US20080070280A1
公开(公告)日：2008-03-20
申请号：US11975799
申请日：2007-10-22
申请人： Bernhard Schilling , Scott Gangloff , Dharti Kothari , Kirk Leister , Linda Matlock , Stephen Zegarelli , Christoph Joosten , Jonathan Basch , Sivakesava Sakhamuri , Steven Lee
发明人： Bernhard Schilling , Scott Gangloff , Dharti Kothari , Kirk Leister , Linda Matlock , Stephen Zegarelli , Christoph Joosten , Jonathan Basch , Sivakesava Sakhamuri , Steven Lee
IPC分类号： C12P21/04
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 inoculation. 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.

3. 发明授权

US07332303B2 Product quality enhancement in mammalian cell culture processes for protein production 有权
标题翻译：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：US07332303B2
公开(公告)日：2008-02-19
申请号：US10740645
申请日：2003-12-18
申请人： Bernhard M. Schilling , Scott Gangloff , Dharti Kothari , Kirk Leister , Linda Matlock , Stephen G. Zegarelli , Christoph E. Joosten , Jonathan D. Basch , Sivakesava Sakhamuri , Steven S. Lee
发明人： Bernhard M. Schilling , Scott Gangloff , Dharti Kothari , Kirk Leister , Linda Matlock , Stephen G. Zegarelli , Christoph E. Joosten , Jonathan D. Basch , Sivakesava Sakhamuri , Steven S. Lee
IPC分类号： C12N15/09 , C12N15/06 , C12N15/56 , A61K38/00
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。

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