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    • 4. 发明申请
    • METHODS FOR PRODUCING RECOMBINANT PROTEINS
    • 生产重组蛋白的方法
    • WO2005024000A1
    • 2005-03-17
    • PCT/GB2004/003747
    • 2004-09-02
    • CELLTECH R & D LIMITEDGLOVER, David, JohnSEHDEV, MukeshREEKS, Dominic, Gambier
    • GLOVER, David, JohnSEHDEV, MukeshREEKS, Dominic, Gambier
    • C12N1/21
    • C12N1/20C12P21/02
    • The present invention provides a method for controlling the partitioning of a recombinant protein between the supernatant and the periplasm in E.coli host cell cultures wherein expression of the recombinant protein by said cells is under the control of an inducible system, which method comprises: a) providing an E.coli host cell culture b) changing the growth rate of the E.coli host cells c) inducing expression of the recombinant protein wherein steps (b) and (c) can be performed in any order or simultaneously; and subsequently d) determining the yield of recombinant protein in the culture supernatant and the E.coli host cell periplasm e) comparing the yield determined in step (d) with the yield determined when at least one other growth rate has been used in step (b) f) selecting a growth rate from the comparison made in step (e) in which the partitioning of the recombinant protein between the supernatant and the periplasm is most suited to the primary recovery of the recombinant protein.
    • 本发明提供了一种用于控制大肠杆菌宿主细胞培养物中上清液和周质之间的重组蛋白质分配的方法,其中所述细胞的重组蛋白质的表达受诱导系统的控制,该方法包括: )提供大肠杆菌宿主细胞培养物b)改变大肠杆菌宿主细胞的生长速率c)诱导重组蛋白质的表达,其中步骤(b)和(c)可以以任何顺序或同时进行; 并且随后d)确定培养上清液和大肠杆菌宿主细胞周质中重组蛋白质的产量e)将步骤(d)中测定的产量与在步骤中至少一种其他生长速率使用时测定的产量进行比较( b)f)从步骤(e)中进行的比较中选择生长速率,其中上清液和周质之间的重组蛋白质的分配最适合于重组蛋白质的初级回收。