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

Subject Area Biological Process Engineering
Term from 2017 to 2023
Project identifier Deutsche Forschungsgemeinschaft (DFG) - Project number 346772917
 
Final Report Year 2024

Final Report Abstract

The benefits of continuous cultivation and production processes have been widely demonstrated by a variety of biopharmaceutical industries in recent years. With the increasing demand for biopharmaceuticals, higher productivity and ever-increasing pressure to reduce manufacturing costs, the biotech industry is showing a continued interest in the development of continuous production systems. Although there are emerging continuous bioprocess technologies on the market, there are almost no actual examples of successful implementation of a fully continuous process for biological products. To accelerate process optimization, it is beneficial to implement the bioreactor with appropriate sensors and technologies for continuous monitoring and control of critical cultivation parameters. In addition, the market is looking for technologies that enable flexible production of individual target proteins and parallelization of production processes. To further advance these development processes, this project proposed the development of an innovative, controlled and continuous cell transfection and cultivation system for recombinant protein production and simultaneous monitoring (e.g., of product and contamination) by integrating various functional microfluidic lab-on-a-chip (LOC) systems. LOC systems offer many advantages for bioprocesses, such as low sample volume, defined and reproducible working conditions, portability, point-of-use diagnostics and the potential to reduce process costs. The main challenges for continuous bioreactor cultivation are the development and integration of functional units and monitoring systems that are suitable for both upstream and downstream processes. Therefore, the focus of this project was on the development of microfluidic systems and novel biosensors that can be integrated directly into a bioprocess. As an important part of the upstream process, a LOC system for continuous and controlled transient gene transfer into host cells was developed and used directly in the cell culture process within this project, enabling flexible production of recombinant proteins. In addition, novel photonic silicon-based and aptamer-based biosensors were developed that can be used for rapid monitoring of target proteins and early detection of potential microbial contamination in cell culture systems. Some of the biosensors developed have also been successfully used for biomarker detection in the field of point-of-care diagnostics. The results of this research project successfully demonstrated the great potential of high-resolution 3D printing technology using biocompatible materials for flexible system development, system miniaturization and integration as well as for individual applications in the field of bioprocess and cell culture technology.

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