US 12,448,456 B2
Method for producing spesolimab
Thomas Wucherpfennig, Ingelheim am Rhein (DE); and Rico Lippmann, Ingelheim am Rhein (DE)
Assigned to Boehringer Ingelheim International GmbH, Ingelheim am Rhein (DE)
Filed by Boehringer Ingelheim International GmbH, Ingelheim am Rhein (DE)
Filed on May 2, 2022, as Appl. No. 17/734,417.
Claims priority of application No. 21171881 (EP), filed on May 3, 2021.
Prior Publication US 2022/0356258 A1, Nov. 10, 2022
Int. Cl. C07K 16/28 (2006.01)
CPC C07K 16/2866 (2013.01) [C07K 2317/14 (2013.01); C07K 2317/24 (2013.01); C07K 2317/41 (2013.01)] 10 Claims
 
1. A method for producing the antibody spesolimab in cell culture comprising
(a) cultivating CHO cells comprising a nucleic acid encoding the antibody spesolimab in a serum-free cell culture medium using fed-batch culture in final N-stage, comprising
(i) seeding the cells in a culture medium, wherein the culture medium is a basal medium, and
(ii) culturing the cells in the culture medium under conditions that allow production of the antibody spesolimab in the cell culture comprising feeding the cells in the cell culture with a feed medium,
wherein Cu2+ is added at 0.35-1.2 μM and iron at 1500 μM or more to the culture medium in step (i), with the basal medium in step (i) or within 2 days after seeding, or a combination thereof;
(b) harvesting the cell culture supernatant comprising the antibody spesolimab; and
(c) purifying the antibody spesolimab from the cell culture supernatant;
wherein an increased iron concentration and a decreased copper concentration added to the culture medium results in the production of the antibody spesolimab having reduced % basic peak group (% BPG) relative to when the antibody spesolimab is produced by the same method using iron concentrations below 1500 μM and copper concentrations above 1.2 μM, wherein % BPG refers to % relative peak area of total sum peak area as determined by cation exchange chromatography (CEX) in an HPLC chromatogram of the spesolimab profile.