Designing a fully automated multi-bioreactor plant for fast DoE optimization of pharmaceutical protein production
Designing a fully automated multi-bioreactor plant for fast DoE optimization of pharmaceutical protein production
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DOI：
10.1002/biot.201200190
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发表时间：
2013-06-01
影响因子：
4.7
通讯作者：
Luttmann, Reiner
中科院分区：
文献类型：
--
作者：
Fricke, Jens;Pohlmann, Kristof;Luttmann, Reiner
为最先进的药物蛋白生产确定最佳表达条件是一个非常耗时和昂贵的过程。在本报告中，描述了在内部设计的小型BIOSTAT (R)多生物反应器工厂中快速和可重复地优化蛋白质表达的方法。新开发的BioPAT (R) MFCS/win实验设计（DoE）模块（德国Sartorius Stedim Systems）将过程控制系统MFCS/win和DoE软件MODDE (R)（瑞典Umetrics AB）连接起来，从而实现全自动优化程序。作为概念的证明，一种商业化的毕赤酵母菌株KM71H已被转化为表达潜在的疟疾疫苗。由于DoE优化程序与初始培养结果相比，该方法使完整蛋白分泌效率提高了一倍。在下一步中，围绕确定的最优值证明了关于过程参数变异性敏感性的鲁棒性。因此，建立了在7个24小时培养周期内显着改善的药物生产工艺。具体来说，根据美国食品和药物管理局（FDA）过程分析技术（PAT）计划中指出的监管要求，将高度仪器化、全自动的多生物反应器平台与适当的培养策略和扩展的DoE软件解决方案相结合，为药用蛋白生产带来了有希望的好处和机会。
The identification of optimal expression conditions for state-of-the-art production of pharmaceutical proteins is a very time-consuming and expensive process. In this report a method for rapid and reproducible optimization of protein expression in an in-house designed small-scale BIOSTAT (R) multi-bioreactor plant is described. A newly developed BioPAT (R) MFCS/win Design of Experiments (DoE) module (Sartorius Stedim Systems, Germany) connects the process control system MFCS/win and the DoE software MODDE (R) (Umetrics AB, Sweden) and enables therefore the implementation of fully automated optimization procedures. As a proof of concept, a commercial Pichia pastoris strain KM71H has been transformed for the expression of potential malaria vaccines. This approach has allowed a doubling of intact protein secretion productivity due to the DoE optimization procedure compared to initial cultivation results. In a next step, robustness regarding the sensitivity to process parameter variability has been proven around the determined optimum. Thereby, a pharmaceutical production process that is significantly improved within seven 24-hour cultivation cycles was established. Specifically, regarding the regulatory demands pointed out in the process analytical technology (PAT) initiative of the United States Food and Drug Administration (FDA), the combination of a highly instrumented, fully automated multi-bioreactor platform with proper cultivation strategies and extended DoE software solutions opens up promising benefits and opportunities for pharmaceutical protein production.