通过添加羧肽酶 B (CpB) 处理步骤，提高细胞培养过程的生产力和电荷变体调节,Biotechnology and Bioengineering

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通过添加羧肽酶 B (CpB) 处理步骤，提高细胞培养过程的生产力和电荷变体调节
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2021-02-24 , DOI: 10.1002/bit.27723
Jianlin Xu ₁ , Shun Zheng ₁ , Zeinab Dawood ₁ , Charles Hill ₁ , Weixin Jin ₁ , Xuankuo Xu ₁ , Julia Ding ₁ , Michael C Borys ₁ , Sanchayita Ghose ₁ , Zheng Jian Li ₁ , Girish Pendse ₂

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细胞培养过程强化的目标是提高生产力，同时保持可接受的质量属性。在本报告中，开发了四种工艺，即传统制造工艺 A，以及通过富集 N-1 种子（工艺 B）、通过灌注 N-1 种子（工艺 C）和通过灌注生产（工艺 D）强化的工艺，用于单克隆抗体的生产。三个强化工艺显着提高了生产率，但是，产品要么未能满足工艺 D 的电荷变体（主峰）规格，要么生产过程需要提前收获以满足第 10 天或第 6 天的电荷变体规格分别用于进程 B 和 C。强化过程的较低主峰是由于较高的 C 末端赖氨酸导致较高的碱性物质。为了解决这个产品质量问题，我们开发了一种酶处理方法，通过引入羧肽酶 B (CpB) 来剪切 C 端赖氨酸，从而显着增加主峰，并为所有强化过程提供可接受且更均匀的产品质量。此外，采用 CpB 处理的工艺 B 和 C 将生物反应器的持续时间延长至第 14 天，滴度分别提高了 38% 和 108%。这种简单而有效的酶处理策略可适用于具有类似产品质量问题的其他工艺。采用 CpB 处理的工艺 B 和 C 将生物反应器的持续时间延长至第 14 天，滴度分别提高了 38% 和 108%。这种简单而有效的酶处理策略可适用于具有类似产品质量问题的其他工艺。采用 CpB 处理的工艺 B 和 C 将生物反应器的持续时间延长至第 14 天，滴度分别提高了 38% 和 108%。这种简单而有效的酶处理策略可适用于具有类似产品质量问题的其他工艺。

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Productivity improvement and charge variant modulation for intensified cell culture processes by adding a carboxypeptidase B (CpB) treatment step

The goal of cell culture process intensification is to improve productivity while maintaining acceptable quality attributes. In this report, four processes, namely a conventional manufacturing Process A, and processes intensified by enriched N-1 seed (Process B), by perfusion N-1 seed (Process C), and by perfusion production (Process D) were developed for the production of a monoclonal antibody. The three intensified processes substantially improved productivity, however, the product either failed to meet the specification for charge variant species (main peak) for Process D or the production process required early harvest to meet the specification for charge variant species, Day 10 or Day 6 for Processes B and C, respectively. The lower main peak for the intensified processes was due to higher basic species resulting from higher C-terminal lysine. To resolve this product quality issue, we developed an enzyme treatment method by introducing carboxypeptidase B (CpB) to clip the C-terminal lysine, leading to significantly increased main peak and an acceptable and more homogenous product quality for all the intensified processes. Additionally, Processes B and C with CpB treatment extended bioreactor durations to Day 14 increasing titer by 38% and 108%, respectively. This simple yet effective enzyme treatment strategy could be applicable to other processes that have similar product quality issues.

更新日期：2021-02-24

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