US 12,331,341 B2
Serum-free cell culture medium
Shadia Abike Oshodi, Jersey City, NJ (US); Amy S. Johnson, Briarcliff Manor, NY (US); and Shawn M. Lawrence, Nyack, NY (US)
Assigned to Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed by Regeneron Pharmaceuticals, Inc., Tarrytown, NY (US)
Filed on Apr. 4, 2022, as Appl. No. 17/712,697.
Application 17/712,697 is a continuation of application No. 14/211,245, filed on Mar. 14, 2014, abandoned.
Claims priority of provisional application 61/790,136, filed on Mar. 15, 2013.
Prior Publication US 2022/0228187 A1, Jul. 21, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/00 (2006.01); C07K 16/18 (2006.01); C12N 5/071 (2010.01); C12P 21/00 (2006.01)
CPC C12P 21/00 (2013.01) [C07K 16/18 (2013.01); C12N 5/0031 (2013.01); C12N 5/0037 (2013.01); C12N 5/0043 (2013.01); C12N 5/005 (2013.01); C12N 5/0682 (2013.01); C12N 2500/22 (2013.01); C12N 2500/24 (2013.01); C12N 2500/32 (2013.01); C12N 2500/34 (2013.01); C12N 2500/46 (2013.01); C12N 2510/02 (2013.01); C12N 2511/00 (2013.01)] 14 Claims
 
1. A method for cultivating CHO cells, comprising the steps of: (a) providing a cell culture medium which is serum-free, comprising 0.6±0.09 mM ornithine and 0.714±0.11 mM putrescine, and (b) propagating or maintaining the CHO cells in the cell culture medium to form a cell culture, wherein the cells express aflibercept, wherein the cell culture has a greater viable cell culture density than an equivalent aflibercept-expressing CHO cell culture grown in a cell culture medium that is supplemented with no more than 2.5 μM putrescine and no ornithine.