US 11,987,638 B1
Prevention of disulfide bond reduction during recombinant production of polypeptides
Yung-Hsiang Kao, San Mateo, CA (US); Michael W. Laird, San Ramon, CA (US); Melody Trexler Schmidt, Danville, CA (US); Rita L. Wong, Redwood City, CA (US); and Daniel P. Hewitt, Sunnyvale, CA (US)
Assigned to Genentech, Inc., South San Francisco, CA (US)
Filed by Genentech, Inc., South San Francisco, CA (US)
Filed on Jan. 11, 2024, as Appl. No. 18/410,949.
Application 18/410,949 is a division of application No. 18/194,471, filed on Mar. 31, 2023.
Application 15/488,917 is a division of application No. 14/043,758, filed on Oct. 1, 2013, abandoned.
Application 14/043,758 is a division of application No. 13/354,223, filed on Jan. 19, 2012, granted, now 8,574,869, issued on Nov. 5, 2013.
Application 18/194,471 is a continuation of application No. 17/087,313, filed on Nov. 2, 2020, granted, now 11,639,395, issued on May 2, 2023.
Application 17/087,313 is a continuation of application No. 16/847,317, filed on Apr. 13, 2020, granted, now 10,906,986, issued on Feb. 2, 2021.
Application 16/847,317 is a continuation of application No. 16/240,592, filed on Jan. 4, 2019, granted, now 10,759,866, issued on Sep. 1, 2020.
Application 16/240,592 is a continuation of application No. 15/488,917, filed on Apr. 17, 2017, abandoned.
Application 13/354,223 is a continuation of application No. 12/217,745, filed on Jul. 8, 2008, abandoned.
Claims priority of provisional application 60/948,677, filed on Jul. 9, 2007.
This patent is subject to a terminal disclaimer.
Int. Cl. C07K 16/28 (2006.01); A61K 39/395 (2006.01); C07K 1/14 (2006.01)
CPC C07K 16/2887 (2013.01) [A61K 39/39591 (2013.01); C07K 1/14 (2013.01); C07K 2317/24 (2013.01)] 30 Claims
 
1. A method for the prevention of the reduction of a disulfide bond in a therapeutic protein comprising an Fe region of an IgG1 antibody expressed in a recombinant Chinese Hamster Ovary (CHO) host cell, comprising supplementing the pre-harvest or harvested culture fluid of said recombinant CHO host cell with a thioredoxin inhibitor,
wherein the thioredoxin inhibitor is selected from the group consisting of:
[i] cupric sulfate added following completion of the cell culture process in a concentration between about 5 mM and about 100 mM,
[ii] 1-fluoro-2,4-dinitrobenzene or dehydroepiandrosterone (DHEA) added in a concentration between about 0.05 mM and about 5 mM, or
[iii] ethylenediamine tetraacetic acid (EDTA) added in a concentration between about 10 mM and about 50 mM.