US 11,952,605 B2
Methods for increasing mannose content of recombinant proteins
Jian Wu, Acton, MA (US); Sean Davern, Seattle, WA (US); Simina Crina Petrovan, Wakefield, MA (US); Michael Charles Brandenstein, Woodinville, WA (US); Katherine Rose Lindahl, Somerville, MA (US); and Shawn Erik Lillie, Puyallup, WA (US)
Assigned to Amgen Inc., Thousand Oaks, CA (US)
Filed by AMGEN INC., Thousand Oaks, CA (US)
Filed on Feb. 22, 2023, as Appl. No. 18/172,670.
Application 18/172,670 is a continuation of application No. 17/704,662, filed on Mar. 25, 2022.
Application 17/704,662 is a continuation of application No. 16/654,933, filed on Oct. 16, 2019, granted, now 11,319,568, issued on May 3, 2022.
Application 16/654,933 is a continuation of application No. 15/652,138, filed on Jul. 17, 2017, abandoned.
Application 15/652,138 is a continuation of application No. 14/776,404, granted, now 9,822,388, issued on Nov. 21, 2017, previously published as PCT/US2014/022738, filed on Mar. 10, 2014.
Claims priority of provisional application 61/784,639, filed on Mar. 14, 2013.
Prior Publication US 2023/0193340 A1, Jun. 22, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 21/00 (2006.01); C07K 16/00 (2006.01); C12N 1/38 (2006.01); C12N 5/00 (2006.01)
CPC C12P 21/005 (2013.01) [C07K 16/00 (2013.01); C12N 1/38 (2013.01); C12N 5/0037 (2013.01); C07K 2317/14 (2013.01); C07K 2317/41 (2013.01); C12N 2500/32 (2013.01); C12N 2500/34 (2013.01); C12N 2510/02 (2013.01)] 26 Claims
 
1. A method of modulating the amount of the mannose-5 glycoform of an Immunoglobulin G2 (IgG2) molecule in an IgG2 composition, wherein said IgG2 composition is produced by a Chinese Hamster Ovary (CHO) cell culture, comprising:
(a) establishing a cell culture of CHO cells in a bioreactor, wherein the CHO cell produces an IgG2 molecule;
(b) limiting the amount of glucose in the bioreactor, wherein the concentration of the glucose in the spent medium of the bioreactor is from about 0 to 3 g/L;
(c) supplementing the cell culture with a feeding medium comprising galactose, such that the concentration of galactose in the resulting spent medium of the bioreactor is above 2.5 g/L; and
(d) harvesting and purifying the IgG2 composition from the cell culture.