The enhancement of antibody concentration and achievement of high cell density CHO cell cultivation by adding nucleoside

Abstract

Recently, with the dramatic increase in demand for therapeutic antibodies, Chinese hamster ovary (CHO) cell culture systems have made significant progress in recombinant antibody production. Over the past two decades, recombinant antibody productivity has been improved by more than 100-fold. Medium optimization has been identified as an important key approach for increasing product concentrations. In this study, we evaluated the effects of deoxyuridine addition to fed-batch cultures of antibody-expressing CHO cell lines. Furthermore, we investigated the effects of combined addition of deoxyuridine, thymidine, and deoxycytidine. Our results suggest that addition of these pyrimidine nucleosides can increase CHO cell growth, with no significant change in the specific production rate. As a result of the increased cell growth, the antibody concentration was elevated and we were able to achieve more than 9 g/L during 16 days of culture. Similar effects of nucleoside addition were observed in fed-batch cultures of a Fab fragment-expressing CHO cell line, and the final Fab fragment concentration was more than 4 g/L. This nucleoside addition strategy could be a powerful platform for efficient antibody production.

Development of an intensified fed-batch production platform with doubled titers using N-1 perfusion seed for cell culture manufacturing

“…Higher titers continue to be reported in recent literature (e.g. 9-10 g/L titer in 14-18 days) (Handlogten et al 2018;Huang et al 2010;Takagi et al 2017), suggesting that potential remains for further increases in productivity and reduction in cost using fed-batch operation.…”

“…Overall, limited literature is available for fed-batch process development with very high (~ 10 g/L) fed-batch titers. Since 10 g/L titer on Day 18 for two proteins was reported for the first time (Huang et al 2010), 9-10 g/L titers were reported for production of one mAb on Day 14 or Day 16 in two other case studies (Handlogten et al 2018;Takagi et al 2017). Conventional SDs of ≤ 1.5 × 10 6 cells/mL were used in the above literature.…”

Supplementation of Nucleosides During Selection can Reduce Sequence Variant Levels in CHO Cells Using GS/MSX Selection System

“…Supplementing the selection media with nucleosides improved nucleotide and DNA synthesis, enhancing cell growth and recovery. Similarly, a recent study has indicated that supplementing nucleosides to the fed‐batch cultures increased CHO cell growth and in turn achieve higher titer …”

Uridine modulates monoclonal antibody charge heterogeneity in Chinese hamster ovary cell fed-batch cultures

“…Chen et al illustrated that hypoxanthine and thymidine can promote initial cell growth and volumetric production of mAb without effects on long-term stability of antibody-producing CHO cells (Chen et al 2011). Deoxyuridine were observed to increase the mAb concentration to 5.5 g/L compared with 3.3 g/L of no addition of deoxyuridine, and there were no significant differences in the specific production rate (Takagi et al 2017). In terms of mAb's quality, it was clarified that feeding of uridine, galactose and manganese chloride can increase the galactosylation level (Gramer et al 2011).…”