| CPC C12N 9/2474 (2013.01) [C12Y 302/01035 (2013.01)] | 15 Claims |
|
1. A method for producing a recombinant protein that comprises at least one of hyaluronidase PH20 or a variant thereof, the method comprising:
first culturing host animal cells expressing the recombinant protein until an integral viable cell density of the host animal cells reaches 20×106 to 120×106 cells×day/mL, wherein first culturing is performed within a first temperature range from 35° C. to 38° C.; and
subsequently second culturing at least part of the host animal cells from first culturing for 2 to 18 days, which produces the recombinant protein, wherein second culturing is performed within a second temperature range from 28° C. to 34° C.,
wherein the second culturing maintains (a) a residual glucose concentration within a range from 0.001 g/L to 4.5 g/L and/or (b) pH within a range from 6.8 to 7.2 in a culture medium therefor,
wherein sialylation of N-glycan of the produced recombinant protein is 1 to 38%, and
wherein the produced recombinant protein has a hyaluronidase enzymatic activity at 10,000 units/mL or higher.
|