The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Gordon Street, London, WC1E 6BT, UK.
Ludger Ltd, Culham Science Centre, Abingdon, Oxfordshire, UK.
Biotechnol J. 2021 Nov;16(11):e2100360. doi: 10.1002/biot.202100360. Epub 2021 Sep 21.
The advancement of microbioreactor technology in recent years has transformed early- and mid-stage process development. The monitoring and control capabilities of microbioreactors not only promote the quick accumulation of process knowledge but has also led to an increased scalability when compared to traditionally used systems such as shake flasks and microtitre plates. This study seeks to establish a framework for the micro-Matrix microbioreactor (Applikon-Biotechnology BV) as process development tool. Using the Dual Indicator System for Mixing Time, the system was initially characterized for mixing properties at varying operating conditions, which was found to yield mixing times between 0.9 and 41.8 s. A matched mixing time was proposed as scale-down criterion for an IgG4 producing GS-CHO fed-batch process between a 5 L stirred tank reactor (STR) and the micro-Matrix microbioreactor. Growth trends, maximum viable cell concentrations, final titre, and glycoprofiles were nearly identical at both scales. The scale-down model was then employed to optimize a bolus feeding regime using response surface methodology, which led to a 25.4% increase of the space-time yield and a 25% increase of the final titre. The optimized feeding strategy was validated at the small-scale and successfully scaled up to the 5 L STR. This work for the first time provides a framework of how the micro-Matrix microbioreactor can be implemented in a bioprocess development workflow and demonstrates scalability of growth and production kinetics as well as IgG4 glycosylation between the micro-Matrix and a benchtop-scale STR system.
近年来，微生物反应器技术的进步改变了早期和中期工艺开发。微生物反应器的监测和控制能力不仅促进了工艺知识的快速积累，而且与摇瓶和微量滴定板等传统使用的系统相比，提高了可扩展性。本研究旨在建立微矩阵微生物反应器（Applikon-Biotechnology BV）作为工艺开发工具的框架。使用混合时间双指示剂系统，对不同操作条件下的混合特性进行了初始表征，结果发现混合时间在 0.9 到 41.8 秒之间。提出了匹配的混合时间作为 IgG4 生产 GS-CHO 分批补料工艺从 5L 搅拌罐反应器 (STR) 到微矩阵微生物反应器的缩小规模标准。在两个规模上，生长趋势、最大活细胞浓度、最终滴度和糖基化谱几乎相同。然后，使用响应面法对该缩小模型进行了优化补料策略的优化，从而使时空产率提高了 25.4%，最终滴度提高了 25%。在小规模上验证了优化的进料策略，并成功扩展到 5L STR。这项工作首次提供了一个框架，说明微矩阵微生物反应器如何可以在生物工艺开发工作流程中实施，并证明了在微矩阵和台式 STR 系统之间生长和生产动力学以及 IgG4 糖基化的可扩展性。
