公开(公告)号：US20140080180A1

公开(公告)日：2014-03-20

申请号：US13826166

申请日：2013-03-14

Applicant: Genentech, Inc.

Inventor： Michael W. Laird ,  Richard St. John ,  Jane V. Gunson ,  Kim Kaleas ,  Deepa Nadarajah ,  Rachel Adams ,  Bradley R. Snedecor

IPC: C12P21/00 ,  C07K16/00

CPC classification number: C12P21/00 ,  C07K16/00

Abstract: 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.

Abstract translation: 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。