US 10,889,790 B2
Growth of bacterial host cells in gas permeable low-density polyethylene bags for production of plasmid DNA or recombinant molecules
Isabelle Louise Chambers, Fargo, ND (US)
Assigned to ALDEVRON, LLC, Fargo, ND (US)
Filed by ALDEVRON, LLC, Fargo, ND (US)
Filed on Mar. 11, 2020, as Appl. No. 16/816,187.
Claims priority of provisional application 62/816,664, filed on Mar. 11, 2019.
Prior Publication US 2020/0291338 A1, Sep. 17, 2020
Int. Cl. C12N 1/20 (2006.01); C12N 1/22 (2006.01); C12N 15/70 (2006.01); C12N 15/74 (2006.01); C12M 1/00 (2006.01); C12M 1/04 (2006.01); C12P 21/00 (2006.01); G01N 1/28 (2006.01)
CPC C12M 23/14 (2013.01) [C12M 23/24 (2013.01); C12N 15/70 (2013.01); C12P 21/00 (2013.01); G01N 1/28 (2013.01)] 20 Claims
1. A method of growing bacterial host cells in a gas-permeable bag comprising the steps of:
providing a starter culture of bacterial host cells, wherein the starter culture comprises at least one colony of the host cells transformed with plasmids comprising an antibiotic resistance gene;
providing the gas-permeable bag;
adding media, an antibiotic, and the starter culture host cells to the gas-permeable bag to form a main culture, wherein the antibiotic is selected such that starter culture host cells expressing the antibiotic resistance gene are viable in the presence of the antibiotic; and
incubating the gas-permeable bag containing the main culture such that host cells grow to a desired concentration.