Development of integrated continuous flow systems for transient transfection, cultivation and monitoring of mammalian cells

开发用于瞬时转染、培养和监测哺乳动物细胞的集成连续流系统

• 批准号: 346772917
• 负责人: Professorin Dr. Janina Bahnemann
• 金额: --
• 依托单位: Institut für Physik
• 依托单位国家: 德国
• 项目类别: Independent Junior Research Groups
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/346772917?language=en
• 关键词: Development; integrated; continuous; flow; systems

Benefits of continuous cultivation and production processes have been widely demonstrated by a variety of manufacturing industries in recent years. With growing demand for biopharmaceuticals, higher productivity and ever-increasing pressure to reduce manufacturing costs, the biotech industry seems to exhibit a persisting interest in the development of continuous bio-manufacturing systems. Although there are emerging continuous bioprocessing technologies available in the market, there are almost no concrete examples for successful implementation of a fully continuous process for biological products. To accelerate the process optimization it is advantageous to implement the bioreactor with appropriate technology for continuous monitoring and regulation of critical cultivation parameters. In addition, the market is seeking for rapid gene delivery technologies that would allow flexible production of individual target proteins and parallelization of production processes. To further enhance this development process, the construction of a novel controlled and continuous cell transfection and cultivation system for recombinant protein production and simultaneous analyte monitoring by integration of different functional microfluidic lab-on-a-chip (LOC) devices is proposed. LOC devices offer many advantages for bioprocesses such as small sample volume, defined and reproducible working conditions, portability, point-of-care (POC) diagnosis, and the potential of reducing process costs. Main challenges for continuous bioreactor cultivation are the development and integration of functional units and monitoring systems, which are suitable for both upstream and downstream processes. Herein, this project will focus on the upstream process and development of new sensors to monitor cell culture systems. As an important part of the upstream process, this project aims to design, fabricate and integrate a LOC for continuous and controlled transient gene transfer to the host cells for recombinant protein production. Furthermore, a novel electromechanical aptamer-based biosensor for rapid monitoring of target proteins as well as for early detection of potential microbial contamination is proposed.

近年来，各种制造业广泛展示了持续耕作和生产过程的好处。随着对生物制药的需求不断增长，生产率不断提高，降低制造成本的压力越来越大，生物技术行业似乎对开发连续生物制造系统表现出了持久的兴趣。尽管市场上有新兴的连续生物加工技术，但几乎没有成功实施生物产品完全连续工艺的具体例子。为了加速工艺优化，采用合适的技术实现生物反应器，对关键培养参数进行连续监测和调节是有利的。此外，市场正在寻求快速基因传递技术，这种技术将允许灵活地生产单个目标蛋白，并实现生产过程的并行化。为了进一步加强这一开发过程，提出了通过集成不同功能的微流控芯片实验室(LOC)设备来构建一种新型的可控和连续的细胞转染和培养系统，用于重组蛋白的生产和同时监测分析物。LOC设备为生物工艺提供了许多优势，例如样品体积小、定义的和可重复的工作条件、便携性、护理点(POC)诊断以及降低工艺成本的潜力。生物反应器连续培养的主要挑战是开发和集成既适用于上游工艺又适用于下游工艺的功能单元和监测系统。在这里，这个项目将集中在上游过程和开发新的传感器来监测细胞培养系统。作为上游过程的重要组成部分，该项目旨在设计、制造和集成一种LOC，用于连续和受控的瞬时基因转移到宿主细胞，以生产重组蛋白。此外，还提出了一种新型的基于机电适配子的生物传感器，用于快速监测目标蛋白以及早期检测潜在的微生物污染。