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    • 2. 发明申请
    • HARVEST OPERATIONS FOR RECOMBINANT PROTEINS
    • 重组蛋白的收获操作
    • US20140080180A1
    • 2014-03-20
    • US13826166
    • 2013-03-14
    • Genentech, Inc.
    • Michael W. LairdRichard St. JohnJane V. GunsonKim KaleasDeepa NadarajahRachel AdamsBradley R. Snedecor
    • C12P21/00C07K16/00
    • C12P21/00C07K16/00
    • The present invention contemplates methods of producing a recombinant protein comprising fermenting a prokaryotic host cell wherein said prokaryotic host cell has been transformed with a nucleic acid encoding said recombinant protein, harvesting said recombinant protein under conditions where dO2 levels are greater than 0%, purifying said recombinant protein to a filtered bulk, wherein said filtered bulk does not contain detectable DHNA-recombinant protein adduct, as measured by an IEC assay at 310 nm. Furthermore, method of producing a recombinant protein comprising fermenting a menE gene-deleted prokaryotic host cell wherein said prokaryotic host cell has been transformed with a nucleic acid encoding said recombinant protein, harvesting said recombinant protein, purifying said recombinant protein to a filtered bulk, wherein said filtered bulk does not contain detectable DHNA-recombinant protein adduct, as measured by an IEC assay at 310 nm, wherein the recombinant protein yield is increased by about 20% or greater is contemplated.
    • 本发明涉及生产包含发酵原核宿主细胞的重组蛋白的方法,其中所述原核宿主细胞已用编码所述重组蛋白的核酸转化,在dO2水平大于0%的条件下收获所述重组蛋白,纯化所述 重组蛋白转化成过滤体积,其中所述过滤的体积不含可检测的DHNA重组蛋白质加合物,如通过IEC测定在310nm测量的。 此外,制备包含发酵menE基因缺失的原核宿主细胞的重组蛋白质的方法,其中所述原核宿主细胞已经用编码所述重组蛋白的核酸转化,收获所述重组蛋白质,将所述重组蛋白质纯化成过滤体积,其中 所述过滤体积不包含可检测的DHNA重组蛋白质加合物,如通过在310nm的IEC测定法测定的,其中重组蛋白质产量提高约20%或更高。