Abstract
Interchain disulfide bonds in monoclonal antibodies may be reduced during large-scale mAb production using Chinese hamster ovary (CHO) cells. This reaction lowers the mAb product yield and purity; however, it may be prevented by screening cell lines that are unsusceptible to reduction and using them in mAb production. Antibody reduction susceptibility may be cell line-dependent. To the best of our knowledge, however, an efficient method of screening reduction-unsusceptible CHO cell lines has not been previously reported. Here, we report a novel screening method that can simultaneously detect and identify mAb reduction susceptibility in lysates containing ≤ 48 CHO cell lines. This evaluation system was equally effective and generated similar results at all culture scales, including 250 mL, 3 L, and 1000 L. Furthermore, we discovered that reduction-susceptible cell lines contained higher total intracellular nicotinamide adenine dinucleotide phosphate (NADPH) and NADP+ concentrations than reduction-unsusceptible cell lines, regardless of whether they expressed immunoglobulin (Ig)G4 or IgG1. NADPH or NADP+ supplementation in the lysate of reduction-unsusceptible cells resulted in mAb reduction. Application of the innovative CHO cell line screening approach could mitigate or prevent reductions in large-scale mAb generation from CHO cells.
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The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
We thank Yuka Mashio for helping with sample preparation and Mayumi Kaniwa and Misaki Furuie for helping with cell culture. We would like to thank Editage (www.editage.com) for English language editing.
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Conceptualization: TY; Methodology: TY; Formal analysis and investigation: TY, MF, YM, TM, SK; Writing—original draft preparation: TY; Writing—review and editing: TY, RN, KY, KW; Supervision: KY, KW.
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Yamaguchi, T., Fukuda, M., Matsumoto, Y. et al. New high-throughput screening method for Chinese hamster ovary cell lines expressing low reduced monoclonal antibody levels: application of a system controlling the gas phase over cell lysates in miniature bioreactors and facilitating multiple sample setup. Cytotechnology 75, 421–433 (2023). https://doi.org/10.1007/s10616-023-00587-x
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DOI: https://doi.org/10.1007/s10616-023-00587-x