Abstract
Continuous production of biologics, a growing trend in the biopharmaceutical industry, requires a reliable and efficient cell retention device that also maintains cell viability. Current filtration methods, such as tangential flow filtration using hollow-fiber membranes, suffer from membrane fouling, leading to significant reliability and productivity issues such as low cell viability, product retention, and an increased contamination risk associated with filter replacement. We introduce a novel cell retention device based on inertial sorting for perfusion culture of suspended mammalian cells. The device was characterized in terms of cell retention capacity, biocompatibility, scalability, and long-term reliability. This technology was demonstrated using a high concentration (>20 million cells/mL) perfusion culture of an IgG1-producing Chinese hamster ovary (CHO) cell line for 18–25 days. The device demonstrated reliable and clog-free cell retention, high IgG1 recovery (>99%) and cell viability (>97%). Lab-scale perfusion cultures (350 mL) were used to demonstrate the technology, which can be scaled-out with parallel devices to enable larger scale operation. The new cell retention device is thus ideal for rapid perfusion process development in a biomanufacturing workflow.
Highlights
In the biopharmaceutical industry, continuous bioprocessing is widely recognized as a generation biomanufacturing platform for reducing manufacturing cost and improving product quality[1, 2]
Alternating Tangential-flow Filtration (ATF) remains susceptible to membrane fouling[8, 10, 16, 17]
Scale-out through parallelization of devices can increase the overall flow throughput further[33,34,35]. Sensitive cells such as mesenchymal stem cells and leukocytes have been tested in the spiral microchannels demonstrating that this processing does not affect indicators of cell viability, such as membrane permeability and surface proteins[36, 37]
Summary
Continuous bioprocessing is widely recognized as a generation biomanufacturing platform for reducing manufacturing cost and improving product quality[1, 2]. Retained behind the hollow-fiber membrane filter in TFF and ATF7, 8, 11, 13, 18, 19 due to membrane fouling and concentration polarization[14, 20] This potentially diminishes protein recovery and increases the protein residence time in the bioreactor. Inertial microfluidics[25,26,27], one of the most successful methods for high-throughput cell sorting[27], utilizes a combination of hydrodynamic forces dependent on particle size in order to focus and separate particles laterally in a continuous flow within the channel. Scale-out through parallelization of devices can increase the overall flow throughput further (up to 1 L/min)[33,34,35] Sensitive cells such as mesenchymal stem cells and leukocytes have been tested in the spiral microchannels demonstrating that this processing does not affect indicators of cell viability, such as membrane permeability and surface proteins[36, 37]. It was shown that spiral cell sorting does not induce up-regulation of a shear stress-related gene of the CHO cells[33]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
