US 12,084,686 B2
Antibodies with modulated glycan profiles
Christopher Kenyon Crowell, Thousand Oaks, CA (US); Jian Wu, Acton, MA (US); Neil Kitchen, Alberta (CA); Alison Jean Gillespie, Seattle, WA (US); Simina Crina Petrovan, Lynnwood, WA (US); and Michael Charles Brandenstein, Woodinville, WA (US)
Assigned to AMGEN INC., Thousand Oaks, CA (US)
Appl. No. 17/051,528
Filed by Amgen Inc., Thousand Oaks, CA (US)
PCT Filed Apr. 30, 2019, PCT No. PCT/US2019/029850
§ 371(c)(1), (2) Date Oct. 29, 2020,
PCT Pub. No. WO2019/213043, PCT Pub. Date Nov. 7, 2019.
Claims priority of provisional application 62/665,045, filed on May 1, 2018.
Prior Publication US 2021/0062156 A1, Mar. 4, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12P 21/00 (2006.01); C07K 16/00 (2006.01); C07K 16/28 (2006.01); C12N 1/38 (2006.01); C12N 5/071 (2010.01); C12N 5/16 (2006.01)
CPC C12N 5/0682 (2013.01) [C07K 16/2875 (2013.01); C07K 2317/14 (2013.01); C07K 2317/92 (2013.01); C12N 2500/34 (2013.01)] 10 Claims
 
1. A method of increasing the level of high-mannose present on denosumab molecules, wherein said denosumab molecules are recombinantly-expressed by a mammalian host cell, comprising:
(a) incubating said mammalian host cell in a first culture medium during growth phase until the cell density is at least 1×106 viable cells/mL, wherein said first culture medium comprises from 1 g/L to 20 g/L glucose; and subsequently
(b) incubating host cells from step (a) in a second culture medium during production phase to express said denosumab molecules, wherein said second culture medium comprises from 0 g/L to 10 g/L glucose and from 5 g/L to 20 g/L galactose;
wherein from 2% to 14% of the denosumab molecules comprise high-mannose glycan at N-298 site.