Investigating the influence of physiologically relevant hydrostatic pressure on CHO cell batch culture

Menglin Shang,Bee Luan Khoo,Jean-Francois P Hamel,Chwee Teck Lim,Taehong Kwon,Jongyoon Han

doi:10.1038/s41598-020-80576-8

Abstract

Chinese hamster ovary (CHO) cells have been the most commonly used mammalian host for large-scale commercial production of therapeutic proteins, such as monoclonal antibodies. Enhancement of productivity of these CHO cells is one of the top priorities in the biopharmaceutical industry to reduce manufacturing cost. Although there are many different methods (e.g. temperature, pH, feed) to improve protein production in CHO cells, the role of physiologically relevant hydrostatic pressure in CHO cell culture has not been reported yet. In this study, four different hydrostatic pressures (0, 30, 60, and 90 mmHg) were applied to batch CHO cells, and their cell growth/metabolism and IgG1 production were examined. Our results indicate that hydrostatic pressure can increase the maximum cell concentration by up to 50%. Moreover, overall IgG1 concentration on Day 5 showed that 30 mmHg pressure can increase IgG1 production by 26%. The percentage of non-disulphide-linked antibody aggregates had no significant change under pressure. Besides, no significant difference was observed between 30 mmHg and no pressure conditions in terms of cell clumping formation. All these findings are important for the optimization of fed-batch or perfusion culture for directing cell growth and improving antibody production.

Highlights

Chinese hamster ovary (CHO) cells have been the most commonly used mammalian host for largescale commercial production of therapeutic proteins, such as monoclonal antibodies
By constructing a pressurized batch culture system, we found that hydrostatic pressure of 30–90 mmHg slowed down initial CHO cell proliferation but increased the maximum cell concentration at the end phase of the culture
We have demonstrated that both cell concentration and antibody production from CHO cell culture can be significantly enhanced by the application of moderate hydrostatic pressure

Summary

Introduction

Chinese hamster ovary (CHO) cells have been the most commonly used mammalian host for largescale commercial production of therapeutic proteins, such as monoclonal antibodies. No significant difference was observed between 30 mmHg and no pressure conditions in terms of cell clumping formation All these findings are important for the optimization of fed-batch or perfusion culture for directing cell growth and improving antibody production. To meet the rapidly growing demand for therapeutic proteins and reduce manufacturing cost, several strategies have been developed to improve biopharmaceutical productions in CHO cells These approaches can be categorized into two groups: cell engineering and bioprocess o ptimization[8]. They found that 0.8 MPa (~ 6000 mmHg above atmosphere pressure) static pressure can enhance human granulocyte–macrophage stimulating factor (hGMCSF) production in adherent CHO cells and lower its intracellular pH from 6.60 to 5.2423–25 All these studies tested pressure of ~ 6000 mmHg, which was significantly higher than the hydrostatic pressure in vivo (typically less than 120 mmHg). At the same osmotic pressure of 450 mOsm/kg, the specific yield of tissue plasminogen activator (tPA) was maximized in suspension CHO cell culture but only reached 50% of its maximal value in adherent c ulture[26]

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Discussion

Conclusion