Improving titer while maintaining quality of final formulated drug substance via optimization of CHO cell culture conditions in low-iron chemically defined media
Improving titer while maintaining quality of final formulated drug substance via optimization of CHO cell culture conditions in low-iron chemically defined media
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通过优化低铁化学成分确定培养基中的 CHO 细胞培养条件，提高效价，同时保持最终配制药物的质量
DOI：
10.1080/19420862.2018.1433978
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发表时间：
2018
期刊：
影响因子：
5.3
通讯作者：
Z. Li
中科院分区：
文献类型：
--
作者：
Jianlin Xu;M. Rehmann;Xuankuo Xu;Chao Huang;Jun Tian;N. Qian;Z. Li
摘要 在生物制药工艺开发过程中，提高效价以降低药物生产成本并提供治疗性蛋白质的可比质量属性非常重要，这有助于确保患者的安全和疗效。我们之前报道过，培养基中相对高的铁浓度会增加滴度，但在早期工艺开发过程中会导致融合蛋白的着色不可接受。最终，使用低铁培养基生产出具有可接受颜色的融合蛋白，但在低铁过程中效价显着下降。在这里，在低铁培养基中长期传代可以显着提高效价，同时在后期工艺开发过程中保持可接受的着色。然而，长期传代也通过显着增加碱性变体而引起蛋白质电荷变体谱的变化。因此，我们系统地研究了培养基成分、种子培养条件和下游加工对生产力和质量属性的影响。我们发现从基础培养基中去除β-磷酸甘油（BGP）可以减少基本变异而不影响效价。因此，我们通过延长种子培养年龄和去除 BGP 来实现后期工艺开发、提高滴度并将质量属性与早期工艺相匹配的目标。该工艺也在 500 L 生物反应器中成功扩大规模。此外，我们证明，与低铁培养物相比，高铁中国仓鼠卵巢细胞培养物中存在更高浓度的活性氧，这表明高铁培养基引起药物物质着色的可能机制。最后，讨论了高铁和长期培养提高效价机制的假设。
ABSTRACT During biopharmaceutical process development, it is important to improve titer to reduce drug manufacturing costs and to deliver comparable quality attributes of therapeutic proteins, which helps to ensure patient safety and efficacy. We previously reported that relative high-iron concentrations in media increased titer, but caused unacceptable coloration of a fusion protein during early-phase process development. Ultimately, the fusion protein with acceptable color was manufactured using low-iron media, but the titer decreased significantly in the low-iron process. Here, long-term passaging in low-iron media is shown to significantly improve titer while maintaining acceptable coloration during late-phase process development. However, the long-term passaging also caused a change in the protein charge variant profile by significantly increasing basic variants. Thus, we systematically studied the effect of media components, seed culture conditions, and downstream processing on productivity and quality attributes. We found that removing β-glycerol phosphate (BGP) from basal media reduced basic variants without affecting titer. Our goals for late-phase process development, improving titer and matching quality attributes to the early-phase process, were thus achieved by prolonging seed culture age and removing BGP. This process was also successfully scaled up in 500-L bioreactors. In addition, we demonstrated that higher concentrations of reactive oxygen species were present in the high-iron Chinese hamster ovary cell cultures compared to that in the low-iron cultures, suggesting a possible mechanism for the drug substance coloration caused by high-iron media. Finally, hypotheses for the mechanisms of titer improvement by both high-iron and long-term culture are discussed.
DOI：
10.1016/s0021-9258(19)39745-5
发表时间：
1990-02
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
A. Sturrock;J. Alexander;Jamie Lamb;Catherine M. Craven;Jerry Kaplan
通讯作者：
A. Sturrock;J. Alexander;Jamie Lamb;Catherine M. Craven;Jerry Kaplan
影响因子：
5
作者：
Beckmann, T. F.;Kraemer, O.;Noll, T.
通讯作者：
Noll, T.