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. 2022 Feb 24;22(5):427-436.
doi: 10.1002/elsc.202100062. eCollection 2022 May.

Stress-induced increase of monoclonal antibody production in CHO cells

Jana Schellenberg  1 , Tamanna Nagraik  1 , Ole Jacob Wohlenberg  1 , Sebastian Ruhl  2 , Janina Bahnemann  1 , Thomas Scheper  1 , Dörte Solle  1
Affiliations

Stress-induced increase of monoclonal antibody production in CHO cells

Jana Schellenberg et al. Eng Life Sci. .

Abstract

Monoclonal antibodies (mAbs) are of great interest to the biopharmaceutical industry due to their widely used application as human therapeutic and diagnostic agents. As such, mAb require to exhibit human-like glycolization patterns. Therefore, recombinant Chinese hamster ovary (CHO) cells are the favored production organisms; many relevant biopharmaceuticals are already produced by this cell type. To optimize the mAb yield in CHO DG44 cells a corelation between stress-induced cell size expansion and increased specific productivity was investigated. CO2 and macronutrient supply of the cells during a 12-day fed-batch cultivation process were tested as stress factors. Shake flasks (500 mL) and a small-scale bioreactor system (15 mL) were used for the cultivation experiments and compared in terms of their effect on cell diameter, integral viable cell concentration (IVCC), and cell-specific productivity. The achieved stress-induced increase in cell-specific productivity of up to 94.94.9%-134.4% correlates to a cell diameter shift of up to 7.34 μm. The highest final product titer of 4 g/L was reached by glucose oversupply during the batch phase of the process.

Keywords: CHO; cell size; cell‐specific productivity; mAb; stress.

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Conflict of interest statement

We confirm that all corresponding authors agree with the submission and publication of this paper and that there is no conflict of interest concerning financial and personal relationships. The manuscript does not contain neither experiments using animals nor human studies. Furthermore, we confirm that the article has not been published previously by any of the authors and is not under consideration for publication elsewhere at the time of submission.

Figures

FIGURE 1
FIGURE 1
Overview of this work experimental structure
FIGURE 2
FIGURE 2
VCC and product concentration (mAB) for SF_CO2 with CO2 limitation and SF_Nu with nutrient limitations compared to the SF_Reference (A). Glucose concentration and average cell diameter for CO2 and nutrient limitation compared to the SF_Reference (B). Calculated values for overall average cell diameter, max. difference in cell diameter, IVCC, cell‐specific productivity Q P, and specific productivity with regard to product titer and IVCC (C). mAB, monoclonal antibody; IVCC, integral viable cell concentration; VCC, viable cell concentration
FIGURE 3
FIGURE 3
VCC and product concentration (mAb) for pH_0 and pH_5 with pH shifts compared to the pH_Reference (A). Glucose concentration and average cell diameter for pH shifts compared to the pH_Reference (B). Calculated values for overall average cell diameter, max. difference in cell diameter, IVCC, cell‐specific productivity Q P, and specific productivity with regard to product titer and IVCC (C). mAB, monoclonal antibody; IVCC, integral viable cell concentration; VCC, viable cell concentration
FIGURE 4
FIGURE 4
VCC and product concentration (mAb) for Nu_1.1, Nu_1.2, Nu_2, and Nu_3 with different PM formulations and feeding strategies compared to the Reference in (A). Glucose concentration and cell diameter with different PM formulations and feeding strategies compared to the Reference in (B). Calculated values for average cell diameter, max. difference in cell diameter, IVCC, cell‐specific productivity Q P, and specific productivity with regard to product titer and IVCC are shown in (C). mAB, monoclonal antibody; IVCC, integral viable cell concentration; PM, production media; VCC, viable cell concentration
FIGURE 5
FIGURE 5
Average cell‐specific productivity (avg Q P) plotted against avg. cell diameter for threefold shake flask and pH experiments in the Ambr 15 bioreactor system are shown in (A). Avg Q P plotted against avg. cell diameter for twofold media screening experiments in the Ambr 15 bioreactor system in (B)
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