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    • 2. 发明授权
    • Fermentation process
    • 发酵过程
    • US09346861B2
    • 2016-05-24
    • US14111223
    • 2012-04-12
    • Philippe Marc Helene DehottayPhilippe Goffin
    • Philippe Marc Helene DehottayPhilippe Goffin
    • A61K39/02C12P1/00C12P21/06C07K14/34C07K14/195C12P21/02A61K39/00
    • C07K14/34A61K2039/55544C07K14/195C12P21/02
    • The present invention provides a process for periplasmic expression of a bacterial toxoid comprising the steps of: a) growing a culture of a gram negative host cell in a fermentation medium, wherein the host cell is transformed with a polynucleotide, and wherein the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence; or providing a gram negative host cell wherein the host cell is transformed with a polynucleotide, the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence and wherein the gram negative host cell comprises the bacterial toxoid expressed in the periplasm; a(i)) inducing expression of the bacterial toxoid; b) maturing the host cell, wherein the maturing step comprises: I) subjecting the host cell to a pH shock; II) incubating the host cell with no feed addition; and/or III) subjecting the host cell to a temperature below −20° C.; and c) extracting the bacterial toxoid from the host cell wherein the extraction process comprises osmotic shock.
    • 本发明提供了一种细菌类毒素的周质表达方法,包括以下步骤:a)在发酵培养基中培养革兰氏阴性宿主细胞的培养物,其中用多核苷酸转化宿主细胞,其中多核苷酸编码 细菌类毒素和周质信号序列; 或提供革兰氏阴性宿主细胞,其中宿主细胞用多核苷酸转化,所述多核苷酸编码细菌类毒素和周质信号序列,其中革兰氏阴性宿主细胞包含在周质中表达的细菌类毒素; (i))诱导细菌类毒素的表达; b)使宿主细胞成熟,其中熟化步骤包括:I)使宿主细胞经受pH震荡; II)不加入饲料孵育宿主细胞; 和/或III)使宿主细胞温度低于-20℃; 和c)从宿主细胞中提取细菌类毒素,其中提取过程包括渗透压休克。
    • 4. 发明申请
    • FERMENTATION PROCESS
    • 发酵过程
    • US20140050758A1
    • 2014-02-20
    • US14111223
    • 2012-04-12
    • Philippe Marc Helene DohottayPhilippe Goffin
    • Philippe Marc Helene DohottayPhilippe Goffin
    • C07K14/34
    • C07K14/34A61K2039/55544C07K14/195C12P21/02
    • The present invention provides a process for periplasmic expression of a bacterial toxoid comprising the steps of:a), growing a culture of a gram negative host cell in a fermentation medium, wherein the host cell is transformed with a polynucleotide, and wherein the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence; or providing a gram negative host cell wherein the host cell is transformed with a polynucleotide, the polynucleotide encodes the bacterial toxoid and a periplasmic signal sequence and wherein the gram negative host cell comprises the bacterial toxoid expressed in the periplasm;a(i)) inducing expression of the bacterial toxoid;b) maturing the host ceil, wherein the maturing step comprises;I) subjecting the host cell to a pH shock;II) incubating the host ceil with no feed addition; and/orIII) subjecting the host ceil to a temperature below −20° C.; andc) extracting the bacterial toxoid from the host cell wherein the extraction process comprises osmotic shock.
    • 本发明提供了一种细菌类毒素的周质表达方法,包括以下步骤:a)在发酵培养基中生长革兰氏阴性宿主细胞的培养物,其中宿主细胞用多核苷酸转化,并且其中多核苷酸编码 细菌类毒素和周质信号序列; 或提供革兰氏阴性宿主细胞,其中宿主细胞用多核苷酸转化,所述多核苷酸编码细菌类毒素和周质信号序列,其中革兰氏阴性宿主细胞包含在周质中表达的细菌类毒素; (i))诱导细菌类毒素的表达; b)熟化宿主细胞,其中熟化步骤包括: I)使宿主细胞受到pH震荡; II)不加入饲料孵育宿主细胞; 和/或III)使宿主细胞温度低于-20℃; 和c)从宿主细胞中提取细菌类毒素,其中提取过程包括渗透压休克。