Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Abstract

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.

Mechanisms driving the lactate switch in Chinese hamster ovary cells

“…Galactose (Altamirano, Cairo, & Godia, 2001;Altamirano et al, 2000;Altamirano, Illanes, Becerra, Cairo, & Godia, 2006;Altamirano, Paredes, Illanes, Cairo, & Godia, 2004;Sun et al, 2013;Xu, Hoshan, & Chen, 2016), fructose (Altamirano et al, 2000;Le et al, 2013;Xu et al, 2016), mannose (Altamirano et al, 2000;Berrios, Altamirano, Osses, & Gonzalez, 2011;Xu et al, 2016), and maltose have all been investigated as alternative energy supplies. Altamirano et al (2000) extensively studied the use of galactose in CHO cell cultures.…”

“…Copper is a common example of this (Luo et al, 2012;Xu et al, 2016;Yuk et al, 2015;Yuk et al, 2014). Copper is a common example of this (Luo et al, 2012;Xu et al, 2016;Yuk et al, 2015;Yuk et al, 2014).…”

“…For example, sugar composition can have a large effect on glycosylation of the final product (Berrios et al, 2011;Surve & Gadgil, 2015;Xu et al, 2016) while high copper concentrations were shown to increase basic variants Yuk et al, 2015) and cause product aggregation (Qian et al, 2011). For example, sugar composition can have a large effect on glycosylation of the final product (Berrios et al, 2011;Surve & Gadgil, 2015;Xu et al, 2016) while high copper concentrations were shown to increase basic variants Yuk et al, 2015) and cause product aggregation (Qian et al, 2011).…”

Perfusion cultures require optimum respiratory ATP supply to maximize cell‐specific and volumetric productivities

“…In addition to their economic feasibility (Croughan, Konstantinov, & Cooney, ), continuous processes using cell retention can provide higher cell densities, prolonged cultivation times, both removal of inhibitory by‐products, and continuous substrate addition (Bielser, Wolf, Souquet, Broly, & Morbidelli, ; Voisard, Meuwly, Ruffieux, Baer, & Kadouri, ). Alternating tangential flow (ATF) and tangential flow filtration (TFF) using filtration via hollow fiber modules can potentially reach cell densities of well above 100 × 10 6 cells/ml (Clincke, Mölleryd, Zhang, et al, ) and stable long‐term operation with high viabilities (Warikoo et al, ; Xu, Hoshan, & Chen, ). Nevertheless, additionally maintaining high cell‐specific productivities during operation is crucial to maximize volumetric productivities and facilitate downstream processing (Bausch, Schultheiss, & Sieck, ; Steinebach et al, ).…”

Development of a shake tube‐based scale‐down model for perfusion cultures

“…Recently, mammalian cell perfusion cultures have regained attraction as an alternative to traditional fed‐batch processes for the production of therapeutic proteins, like monoclonal antibodies (mAbs; Clincke et al, ; Karst, Serra, Villiger, Soos, & Morbidelli, ). Perfusion cultures have been used for the production of unstable proteins (Bödeker, Newcomb, Yuan, Braufman, & Kelsey, ; Warikoo et al, ), and are in general a promising tool for the intensification of protein production processes (Hiller, Ovalle, Gagnon, Curran, & Wang, ; Moulijn, Stankiewicz, Grievink, & Górak, ; Subramanian, ; Xu, Hoshan, & Chen, ; Yang, Minkler, Kshirsagar, & Ryll, ). Particular interest has been directed to their application in the frame of continuous end‐to‐end integrated production processes (Godawat, Konstantinov, Rohani, & Warikoo, ; Karst, Scibona, et al, 2017; Karst, Steinebach, & Morbidelli, ; Karst, Steinebach, Soos, & Morbidelli, ; Steinebach et al, ; Warikoo et al, ).…”