Walther Julia, Ribeiro da Costa Tiago, Winkler Lydia, Schaub Jochen, Habicher Tobias
Bioprocess Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riß, Germany.
Front Bioeng Biotechnol. 2025 Jul 18;13:1608889. doi: 10.3389/fbioe.2025.1608889. eCollection 2025.
The production of monoclonal antibodies (mAbs) using Chinese Hamster Ovary (CHO) cell host systems often faces challenges in terms of manufacturing costs and efficiency. To address these challenges, process intensification with high seeding density production processes applying N-1 perfusion is utilized. This study delves into the impact of cell specific perfusion rate (CSPR) and the total exchanged medium in relation to the reactor working volume (iVVD) on cell growth dynamics and metabolic stability in N-1 perfusion cultivations. The effect of varying the perfusion rate increase (PRI) while keeping the amount of exchanged medium constant is investigated, revealing a positive correlation between iVVD and overall cell growth. However, this effect plateaus at higher values, indicating diminishing returns on cell growth with increased medium use. We found that CSPR directly influences the specific metabolic rates of several metabolites and amino acids, accelerating overall metabolism without necessarily affecting growth. Interestingly, the specific metabolic rates are driven by the CSPR after a metabolic adaptation until day 2.5. Besides adjusting perfusion rates every 24 h, the potential benefits of real-time CSPR control using a capacitance probe are explored. While real-time control offers more precise regulation of the perfusion rate, growth and metabolic behavior is comparable to predefined rates within the tested range. This study demonstrates that optimization of factors such as CSPR, iVVD, and PRI can lead to improved cell growth and viability with the potential to decrease media expenditure, thereby reducing manufacturing costs for the production of mAbs using CHO cell host systems.
使用中国仓鼠卵巢（CHO）细胞宿主系统生产单克隆抗体（mAb）在制造成本和效率方面常常面临挑战。为应对这些挑战，采用了应用N-1灌注的高接种密度生产工艺来强化生产过程。本研究深入探讨了细胞比灌注速率（CSPR）以及与反应器工作体积相关的总交换培养基（iVVD）对N-1灌注培养中细胞生长动力学和代谢稳定性的影响。研究了在保持交换培养基量恒定的同时改变灌注速率增加量（PRI）的效果，结果表明iVVD与总体细胞生长呈正相关。然而，这种效果在较高值时趋于平稳，表明随着培养基使用量增加，细胞生长的回报递减。我们发现CSPR直接影响几种代谢物和氨基酸的比代谢速率，加速总体代谢但不一定影响生长。有趣的是，在代谢适应至第2.5天之前，比代谢速率由CSPR驱动。除了每24小时调整一次灌注速率外，还探索了使用电容探头进行实时CSPR控制的潜在益处。虽然实时控制可更精确地调节灌注速率，但在测试范围内，生长和代谢行为与预定义速率相当。本研究表明，优化CSPR、iVVD和PRI等因素可改善细胞生长和活力，有可能减少培养基消耗，从而降低使用CHO细胞宿主系统生产mAb的制造成本。
