Model-based design and control of a small-scale integrated continuous end-to-endmAbplatform
Model-based design and control of a small-scale integrated continuous end-to-endmAbplatform
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DOI：
10.1002/btpr.2995
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发表时间：
2020-04-06
影响因子：
2.9
通讯作者：
Chotteau, Veronique
中科院分区：
文献类型：
--
作者：
Gomis-Fons, Joaquin;Schwarz, Hubert;Chotteau, Veronique
从工艺开发的最早阶段就可获得的连续集成生物工艺允许集成工艺的更容易、更有效和更快的开发和表征以及小规模候选药物的生产。本文中介绍的工艺是基于200 ml交替切向流过滤灌注生物反应器的非常小规模的连续端到端单克隆抗体生产平台的概念验证，该平台与具有基于模型的设计和控制的纯化工艺集成。在一个单一的层析系统中完成了包括蛋白A周期性双柱捕获、病毒灭活、CEX柱和AEX柱的下游过程，纯化时间小于4 h，在CHO细胞的高细胞密度灌注培养中生产了17 d的单克隆抗体，滴度高达1.0 mg/ml。通过对所有色谱步骤进行建模，创建了下游工艺的数字孪生模型。这些模型用于实时决策，通过实施控制策略来自动化和优化过程的操作。通过工艺的稳态操作确保纯化产物的糖基化模式一致。关于杂质的去除，实现了HCP水平至少4个对数的降低。回收率可达60%，纯化产物的最大生产能力为0.8mg/ml/天。
A continuous integrated bioprocess available from the earliest stages of process development allows for an easier, more efficient and faster development and characterization of an integrated process as well as production of small-scale drug candidates. The process presented in this article is a proof-of-concept of a continuous end-to-end monoclonal antibody production platform at a very small scale based on a 200 ml alternating tangential flow filtration perfusion bioreactor, integrated with the purification process with a model-based design and control. The downstream process, consisting of a periodic twin-column protein A capture, a virus inactivation, a CEX column and an AEX column, was compactly implemented in a single chromatography system, with a purification time of less than 4 hr. Monoclonal antibodies were produced for 17 days in a high cell density perfusion culture of CHO cells with titers up to 1.0 mg/ml. A digital twin of the downstream process was created by modelling all the chromatography steps. These models were used for real-time decision making by the implementation of control strategies to automatize and optimize the operation of the process. A consistent glycosylation pattern of the purified product was ensured by the steady state operation of the process. Regarding the removal of impurities, at least a 4-log reduction in the HCP levels was achieved. The recovery yield was up to 60%, and a maximum productivity of 0.8 mg/ml/day of purified product was obtained.