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
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..
| Medienart: |
Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:75 |
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| Enthalten in: |
Cytotechnology - 75(2023), 5 vom: 21. Juli, Seite 421-433 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Yamaguchi, Tsuyoshi [VerfasserIn] |
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| Links: |
Volltext [lizenzpflichtig] |
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| Themen: |
Antibody reduction |
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| Anmerkungen: |
© The Author(s), under exclusive licence to Springer Nature B.V. 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. |
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| doi: |
10.1007/s10616-023-00587-x |
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| funding: |
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| Förderinstitution / Projekttitel: |
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OLC2145285407 |
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| 520 | |a 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. | ||
| 650 | 4 | |a Chinese hamster ovary cell | |
| 650 | 4 | |a Therapeutic monoclonal antibody | |
| 650 | 4 | |a Antibody reduction | |
| 650 | 4 | |a High-throughput cell line screening method | |
| 650 | 4 | |a Total intracellular NADPH and NADP | |
| 650 | 4 | |a concentration | |
| 700 | 1 | |a Fukuda, Mie |4 aut | |
| 700 | 1 | |a Matsumoto, Yuichi |4 aut | |
| 700 | 1 | |a Mori, Takaaki |4 aut | |
| 700 | 1 | |a Kikuchi, Shinsuke |4 aut | |
| 700 | 1 | |a Nagano, Ryuma |4 aut | |
| 700 | 1 | |a Yamamoto, Koichi |4 aut | |
| 700 | 1 | |a Wakamatsu, Kaori |4 aut | |
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