Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations.
In this study, we discussed the development and optimization of an intensified CHO culture process, highlighting medium and control strategies to improve lactate metabolism. A few strategies, including supplementing glucose with other sugars (fructose, maltose, and galactose), controlling glucose level at <0.2 mM, and supplementing medium with copper sulfate, were found to be effective in reducing lactate accumulation. Among them, copper sulfate supplementation was found to be critical for process optimization when glucose was in excess. When copper sulfate was supplemented in the new process, two-fold increase in cell density (66.5 ± 8.4 × 10(6) cells/mL) and titer (11.9 ± 0.6 g/L) was achieved. Productivity and product quality attributes differences between batch, fed-batch, and concentrated fed-batch cultures were discussed. The importance of process and cell metabolism understanding when adapting the existing process to a new operational mode was demonstrated in the study.
Process Development and Engineering, Biologics and Vaccines, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ, 07033, USA. sen.xu@merck.com.
Sen
S
Xu
Process Development and Engineering, Biologics and Vaccines, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ, 07033, USA.
Linda
L
Hoshan
Process Development and Engineering, Biologics and Vaccines, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ, 07033, USA.
Hao
H
Chen
D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods*; D019327:Copper Sulfate / Q000494:pharmacology*; D006224:Cricetinae; D003412:Cricetulus; D005947:Glucose / Q000494:pharmacology*; D019344:Lactic Acid / Q000096:biosynthesis*
D019344:Lactic Acid; D005947:Glucose; D019327:Copper Sulfate
Alternating tangential flow; Copper sulfate; High cell density; Process intensification; Product quality