Small-Scale Membrane-Less Perfusion Bioreactor System for High-Throughput Cell Line Development and Process Optimization

The aim of this project is to develop a fully automated small-scale (<500 mL) membrane-less perfusion bioreactor system for CHO cells at very high concentration (50–100E+6 cells/mL) using a novel cell retention device based on inertial sorting.

Industry Need

Currently, there is no reliable scale-down model for perfusion culture to be used for cell line development and process optimization. Existing perfusion culture using hollow fiber membranes (ATF or TFF) is too difficult to maintain.

Approach

We aim to develop a fully automated smallscale (<500 mL) membrane-less perfusion bioreactor system for CHO cells at very high concentration (50–100E+6 cells/mL) using a novel cell retention device based on inertial sorting.

Impacts

Fully automated small-scale perfusion culture using a membrane-less cell retention device

Rapid perfusion culture process development with a simple and low cost cell retention device

Value Statement/Outcomes

By implementing a scalable, membrane-less perfusion culture system using microfluidic cell retention technology, organizations can eliminate costly filter replacements and reduce product loss from membrane clogging, achieving >99% product recovery. This approach accelerates process development through rapid generation of high-density culture data (up to 50–100E+6 cells/mL) and supports GMP-compliant workflows that ensure consistent product quality and reliability. It also provides a clear pathway to large-scale perfusion culture (>100 L) without compromising regulatory standards.

Outputs/Deliverables

This project focused on the development of a small-scale membrane-less perfusion culture system that could enable more reliable and efficient cell line and process development than conventional hollow fiber membranes

The project developed a scalable microfluidic cell manipulation platform (>1L/min) [1] for implementing a larger production-scale (>100 L) perfusion culture by providing membrane-less cell retention

Manipulation of ultra-high-density cells by elasto-inertial microfluidics and cascaded spiral retention system

Rapid generation of various culture parameter data in small-scale perfusion culture, which can be used for large-scale production culture

Publications

Jeon, H., Kwon, T., Yoon, J., & Han, J. (2022). Engineering a deformation-free plastic spiral inertial microfluidic system for CHO cell clarification in biomanufacturing. Lab on a Chip, 22(2), 272-285. https://doi.org/10.1039/d1lc00995h

Kwon, T., Choi, K., & Han, J. (2021). Separation of Ultra-High-Density Cell Suspension via Elasto-Inertial Microfluidics. Small, 17(39). https://doi.org/10.1002/smll.202101880

Presentations

Jeon, H., Kwon, T., Yoon, J., & Han, J., Biomanufacturing Scale CHO Cell Clarification Using Hard Plastic Spiral Inertial Microfluidic Device, MicroTAS 2021, Virual, October 20, 2021.