PATFP细胞灌流培养关键过程参数的作用机理和过程放大策略的研究

近年发展起来的细胞灌流培养技术可实现细胞连续培养，达到提高应急病毒疫苗产能，降低成本的目的。我国目前缺乏灌流培养装置装备和细胞分离的中空纤维膜柱材料，尚未开展灌流培养过程中关键过程参数对细胞生长和病毒复制作用机制的基础研究，没有建立灌流培养规模放大的技术策略等。这些装备与基础研究的不足限制了灌流培养技术在我国疫苗产业中的发展。.本研究以自主研发的细胞灌流培养系统（PATFP）与重组腺病毒载体疫苗HEK 293细胞培养生产过程为对象，分析PATFP系统剪切力产生的关键影响因素，建立剪切力评价策略以及细胞生理健康状态评价指标，确定PATFP系统的过程设计空间与规模放大策略；利用多变元分析（MVDA）并结合QbD过程研究理念，解析灌流培养过程中的关键过程参数，并建立稳定的过程参数设计空间，建立灌流培养过程高效研发与规模放大技术。研究结果将为细胞灌流培养过程研发及规模放大提供理论支撑。

Recently, the emergence of perfusion culture technology makes continuous cell culture available. The new technology can improve the production capacity of emergency virus vaccine and reduce manufacture cost. However, in China the equipment for cell perfusion culture and hollow fiber membrane column for cell separation is still not developed. Also no basic research on the mechanism of critical process parameters on cell growth and virus replication in the process of perfusion culture was conducted. Furthermore no technical strategy for process scale-up of perfusion culture was established. The lack of equipment and basic research limits the application of perfusion culture technology in Chinese viral vaccine industry..The proposed study will focus on the process of HEK 293 cell culture using the self-developed perfusion culture technology (PATFP) for the bulk manufacture of adenovirus vector COVID-19 vaccine. The key factors of influencing generation of shear force in PATFP cell culture will be investigated. The assessment method of the impaction by shear force towards the cells in bioreactor, and the evaluation index of cell physiological health status will be established. The design space and scale-up technical strategy of PATFP based cell culture process will be developed. The critical process parameters of perfusion culture will be identified by using multivariate analysis (MVDA) in accordance with QbD concept. The stable design space of process parameters was established, and the efficient R & D and scale-up technology of perfusion culture process was established. The outcome of the proposed study will provide theoretical support for the process development and scale-up of cell perfusion culture, and further promote the application of perfusion culture technology in China.