Commercial Process Development, Genzyme - A Sanofi Company, 45 New York Ave, Framingham, Massachusetts 01701, USA.
Biotechnol Bioeng. 2012 Dec;109(12):3018-29. doi: 10.1002/bit.24584. Epub 2012 Aug 6.
In the current environment of diverse product pipelines, rapidly fluctuating market demands and growing competition from biosimilars, biotechnology companies are increasingly driven to develop innovative solutions for highly flexible and cost-effective manufacturing. To address these challenging demands, integrated continuous processing, comprised of high-density perfusion cell culture and a directly coupled continuous capture step, can be used as a universal biomanufacturing platform. This study reports the first successful demonstration of the integration of a perfusion bioreactor and a four-column periodic counter-current chromatography (PCC) system for the continuous capture of candidate protein therapeutics. Two examples are presented: (1) a monoclonal antibody (model of a stable protein) and (2) a recombinant human enzyme (model of a highly complex, less stable protein). In both cases, high-density perfusion CHO cell cultures were operated at a quasi-steady state of 50-60 × 10(6) cells/mL for more than 60 days, achieving volumetric productivities much higher than current perfusion or fed-batch processes. The directly integrated and automated PCC system ran uninterrupted for 30 days without indications of time-based performance decline. The product quality observed for the continuous capture process was comparable to that for a batch-column operation. Furthermore, the integration of perfusion cell culture and PCC led to a dramatic decrease in the equipment footprint and elimination of several non-value-added unit operations, such as clarification and intermediate hold steps. These findings demonstrate the potential of integrated continuous bioprocessing as a universal platform for the manufacture of various kinds of therapeutic proteins.
在当前多样化的产品管线、快速波动的市场需求以及生物类似药竞争日益激烈的环境下，生物技术公司越来越需要开发创新解决方案，以实现高度灵活和具有成本效益的制造。为了满足这些具有挑战性的需求，集成的连续处理，包括高密度灌注细胞培养和直接偶联的连续捕获步骤，可以用作通用的生物制造平台。本研究首次成功展示了将灌注生物反应器与四柱周期性逆流色谱（PCC）系统集成，用于连续捕获候选蛋白治疗药物。介绍了两个实例：（1）单克隆抗体（稳定蛋白模型）和（2）重组人酶（高度复杂、不太稳定的蛋白模型）。在这两种情况下，高密度灌注 CHO 细胞培养在 50-60×10(6)个细胞/mL 的准稳态下运行超过 60 天，实现了比当前灌注或分批过程更高的体积产率。直接集成和自动化的 PCC 系统连续运行 30 天，没有出现基于时间的性能下降迹象。连续捕获过程中观察到的产品质量与批处理柱操作相当。此外，灌注细胞培养和 PCC 的集成导致设备占地面积显著减少，并消除了几个非增值的单元操作，如澄清和中间储存步骤。这些发现表明集成连续生物加工作为制造各种治疗蛋白的通用平台具有潜力。
