Granted Patent B2

US 12,486,524 · App. 17/572,978 · Granted Dec 2, 2025

Inventors: Christopher Hwang (Cambridge, MA); Timothy Johnson (Cambridge, MA); Jason Walther (Cambridge, MA); Cheng Cheng (Cambridge, MA); Jonathan Wang (Cambridge, MA); Neha Shah (Cambridge, MA); Seul-A Bae (Cambridge, MA)

Assignee: Genzyme Corporation

C12P21/00C07K16/00C12N5/0043C12N5/005C07K2317/14C12N2500/50C12N2510/02

Abstract

Claims (12)

1 . A method of culturing a CHO cell in a liquid culture medium to achieve a viable cell density of greater than 60×10 6 cells/mL, the method comprising:

providing a culturing system comprising a vessel, a liquid culture medium disposed within the vessel, and a sparger disposed within the vessel comprising a plurality of pores configured to dispense gas through the sparger into the liquid culture medium; and

perfusion culturing a CHO cell comprising a recombinant protein-encoding nucleic acid in the culturing system comprising the liquid culture medium comprising an amount of poloxamer-188 and antifoam under conditions and a gas flow rate sufficient to produce the recombinant protein in the system, wherein the ratio of antifoam (g/L) to poloxamer-188 (g/L) is about 1.0% to about 3.0% in the liquid culture medium, and wherein:

(i) the plurality of pores are drilled pores with a pore size of 900 μm to 1.1 mm and the poloxamer-188 concentration is 2.5 g/L to 3.1 g/L;

(ii) the plurality of pores are drilled pores with a pore size of 400 μm to 600 μm and the poloxamer-188 concentration is 3.5 g/L to 4.1 g/L;

(iii) the plurality of pores are drilled pores with a pore size of 1 μm to 250 μm and the poloxamer-188 concentration is 4.3 g/L to 8.0 g/L; or

(iv) the plurality of pores are sintered pores with a pore size of 100 μm and the poloxamer-188 concentration is 5.8 g/L, and

wherein the method results in a viable cell density in the liquid culture medium that is greater than 60×10 6 cells/mL.

2 . The method of claim 1 , wherein the plurality of pores are drilled pores with a pore size of 900 μm to 1.1 mm and the poloxamer-188 concentration is 2.5 g/L to 3.1 g/L.

3 . The method of claim 1 , wherein the plurality of pores are drilled pores with a pore size of 1 μm to 250 μm and the poloxamer-188 concentration is 4.3 g/L to 8.0 g/L.

4 . The method of claim 1 , wherein the plurality of pores are drilled pores with a pore size of 400 μm to 600 μm and the poloxamer-188 concentration is 3.5 g/L to 4.1 g/L.

5 . The method of claim 1 , wherein the plurality of pores are sintered pores with a pore size that is 100 μm and the poloxamer-188 concentration is 5.8 g/L.

Assignments (1)

Continuity (3)

References Cited (181)