Advanced Micro/nanofluidic System for Continuous Bioprocessing
High-quality complex biopharmaceutical products, based on cells and proteins, are revolutionizing modern medicine and advancing treatments for various health conditions. In the biopharmaceutical industry, continuous biomanufacturing has emerged as a prominent technology trend aimed at enhancing the quality of biological products while reducing manufacturing costs.
Our research focuses on introducing innovative technologies for high-throughput microfluidic cell separation [1, 2, 4, 5, 6] and nanofluidic protein quality monitoring [3, 4]. Our novel micro/nanofluidic system [4] enables reliable and efficient microfiltration, as well as robust online monitoring of rapid product quality during continuous biomanufacturing.
This groundbreaking technology addresses the limitations associated with current membrane-based microfiltration and quality monitoring technologies. It overcomes challenges such as filter clogging, low product recovery, manual sample preparation, and off-line analysis. By leveraging our micro/nanofluidic system, biopharmaceutical manufacturers can achieve enhanced efficiency, improved product quality, and streamlined manufacturing processes.
Membrane-less cell retention for perfusion bioreactors
We have developed a novel cell retention device for perfusion culture based on inertial sorting [1, 5, 6]. This membrane-less microfiltration utilizes size-dependent hydrodynamic forces to separate suspended mammalian cells. The device has been extensively evaluated for cell retention efficiency, long-term biocompatibility, and scalability, demonstrating its effectiveness in retainining cells and its compatibility for long-term use. Moreover, we have successfully demonstrated its applicability in long-term and small-scale perfusion cultures, showcasing clog-free cell retention and high product recovery. Our finding offer a promising solution for enhancing the performance and efficiency of bioprrocesses, providing reliable and efficient perfusion culture systems without the limitations of traditional membrane-based filtration methods.
Continuous removal of small nonviable CHO cells from bioreactors
Next, we implemented a high-throughput size-based cell separation technique using inertial sorting to remove small dead cells from bioreactor cultivation [2]. Through the optimization of device parameters, we achieved efficient removal of dead cells, even at high throughputs and concentrations. Our work demonstrated the capability of this approach to effectively remove small dead cells from bioreactor cultures, offering a valuable strategy for improving the overall health and viability of cell-based processes.
Continuous online protein quality monitoring during perfusion culture
Finally, we successfully achieved continuous online monitoring of protein purity in the cell culture supernatant during perfusion culture using a novel nanofluidic filter array [3, 4]. This innovative device, integrated with the microfluidic cell retention system, enabled real-time sample preparation and automated monitoring of protein purity for over a week. By providing a robust online sensing technology, the nanofluidic filter array has the potential to replace conventional offline analytical methods and significantly enhance the efficiency and reliability of protein purity monitoring in continuous biomanufacturing processes.
In summary, our research introduces a novel micro/nanofluidic system for the separation and monitoring of cells and proteins in continuous biomanufacturing [4]. This innovative approach has the potential to enhance the long-term reliability and efficiency of biomanufacturing processes. By addressing key challenges in cell and protein separation, our work contributes to the advancement of continuous biomanufacturing technology and holds promise for future applications.
References:
[1] T. Kwon, H. Prentice, J. D. Oliveira, N. Madziva, M. E. Warkiani, J.-F. P. Hamel, and J. Han, “Microfluidic Cell Retention Device for Perfusion of Mammalian Suspension Culture,” Scientific Reports. 7, 6703 (2017). link
[2] T. Kwon, R. Yao, J.-F. P. Hamel, and J. Han, “Continuous Removal of Small Nonviable Suspended Mammalian Cells and Debris from Bioreactors Using Inertial Microfluidics,” Lab on a Chip, 18, 2826 – 2837 (2018). link
[3] S. H. Ko, D. C., W. Ouyang, T. Kwon, P. Karande, and J. Han, “Nanofluidic device for continuous multiparameter quality assurance of biologics,” Nature Nanotechnology, 12, 804 – 812 (2017). link
[4] T. Kwon, S. H. Ko, J.-F. P. Hamel, and J. Han, “Continuous online protein quality monitoring during perfusion culture production using an integrated micro/nanofluidic system,” Analytical Chemistry, 92, 5267 - 5275 (2020). link
[5] L. Yin, W. Y. Au, C. C. Yu, T. Kwon, Z. Lai, M. Shang, M. E. Warkiani, R. Rosche, C. T. Lim, and J. Han, “Miniature auto‐perfusion bioreactor system with spiral microfluidic cell retention device,” Biotechnology and Bioengineering, 118, 1951 - 1961 (2021). link
[6] T. Kwon, K. Choi, and J. Han, “Separation of Ultra-high-density Cell Suspension via Elasto-inertial Microfluidics,” Small, 17, 2101880 (2021). link