US 12,391,769 B2
Methods of making fucosylated and afucosylated forms of a protein
Shahram Misaghi, Clayton, CA (US); John B. Lowe, San Francisco, CA (US); and Bradley Richard Snedecor, Portola Valley, CA (US)
Assigned to Genentech, Inc., South San Francisco, CA (US)
Filed by Genentech, Inc., South San Francisco, CA (US)
Filed on Sep. 25, 2023, as Appl. No. 18/474,014.
Application 18/474,014 is a continuation of application No. 17/148,456, filed on Jan. 13, 2021, granted, now 11,802,165.
Application 17/148,456 is a continuation of application No. 15/968,214, filed on May 1, 2018, granted, now 10,975,168, issued on Apr. 13, 2021.
Application 15/968,214 is a continuation of application No. PCT/US2016/059922, filed on Nov. 1, 2016.
Claims priority of provisional application 62/338,280, filed on May 18, 2016.
Claims priority of provisional application 62/249,828, filed on Nov. 2, 2015.
Prior Publication US 2024/0158536 A1, May 16, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 16/44 (2006.01); C07K 14/435 (2006.01); G01N 33/68 (2006.01)
CPC C07K 16/44 (2013.01) [C07K 14/435 (2013.01); G01N 33/6854 (2013.01); C07K 2317/41 (2013.01); C07K 2317/52 (2013.01); C07K 2317/732 (2013.01); C07K 2319/00 (2013.01)] 11 Claims
 
1. A method of producing a protein, wherein the protein is produced in fucosylated and afucosylated forms at a predetermined ratio, the method comprising: culturing a host cell engineered to express the protein in a culture medium comprising a fucose source in an amount to produce fucosylated and afucosylated forms of the protein at the predetermined ratio of about 50:1 to about 1:50,
wherein the host cell has no GDP-keto-6-deoxymannose-3,5-epimerase,4-reductase (FX),
wherein the host cell is a FX knockout host cell having FX knocked out in all alleles.