Advertisement

 

 

Rapid protein production from stable CHO cell pools using plasmid vector and the cumate gene-switch.

Rapid protein production from stable CHO cell pools using plasmid vector and the cumate gene-switch.
Author Information (click to view)

Poulain A, Perret S, Malenfant F, Mullick A, Massie B, Durocher Y,


Poulain A, Perret S, Malenfant F, Mullick A, Massie B, Durocher Y, (click to view)

Poulain A, Perret S, Malenfant F, Mullick A, Massie B, Durocher Y,

Advertisement
Share on FacebookTweet about this on TwitterShare on LinkedIn

Journal of biotechnology 2017 06 15255() 16-27 pii 10.1016/j.jbiotec.2017.06.009

Abstract

To rapidly produce large amounts of recombinant proteins, the generation of stable Chinese Hamster Ovary (CHO) cell pools represents a useful alternative to large-scale transient gene expression (TGE). We have developed a cell line (CHO(BRI/rcTA)) allowing the inducible expression of recombinant proteins, based on the cumate gene switch. After the identification of optimal plasmid DNA topology (supercoiled vs linearized plasmid) for PEIpro™ mediated transfection and of optimal conditions for methionine sulfoximine (MSX) selection, we were able to generate CHO(BRI/rcTA) pools producing high levels of recombinant proteins. Volumetric productivities of up to 900mg/L were reproducibly achieved for a Fc fusion protein and up to 350mg/L for an antibody after 14days post-induction in non-optimized fed-batch cultures. In addition, we show that CHO pool volumetric productivities are not affected by a freeze-thaw cycle or following maintenance in culture for over one month in the presence of MSX. Finally, we demonstrate that volumetric protein production with the CR5 cumate-inducible promoter is three- to four-fold higher than with the human CMV or hybrid EF1α-HTLV constitutive promoters. These results suggest that the cumate-inducible CHO(BRI/rcTA) stable pool platform is a powerful and robust system for the rapid production of gram amounts of recombinant proteins.

Submit a Comment

Your email address will not be published. Required fields are marked *

five × 4 =

[ HIDE/SHOW ]