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Science

The bioprocessing industry is increasingly shifting towards continuous downstream processing to capitalize on its advantages such as smaller footprints, shorter cycle times, higher equipment utilization, and improved scalability [2-4]. Existing low pH viral inactivation methods for continuous processing typically rely on predictive models [5-7] to adjust pH levels. However, these methods struggle to manage real-world variations in the processes, such as fluctuations in protein concentration.

Societal Impact

This study introduces an inline viral inactivation system (IVIS) to address these challenges by enabling precise and real-time pH control, ensuring process consistency even in dynamic production environments. Unlike traditional methods, IVIS adapts dynamically to process variations, offering superior accuracy and ease of implementation. In addition, it eliminates the usage of mixer tanks and overcomes the inherent inefficiencies of batch-mode operations, supporting the shift toward continuous processing. This innovation helps ensure consistent product quality across diverse manufacturing environments, making biologics production more efficient and scalable.

Technical Summary

The IVIS integrates:

1. Inline Acidification System: Employs dual-mode adaptive control for precise pH adjustments based on real-time inline sensor readings. This compensates for process variability and batch inconsistencies, especially relevant during development stages.
2. Coiled Flow Inversion Reactor: Maintains controlled incubation for acidified protein eluates, ensuring a consistent and narrow residence time distribution critical for viral inactivation.

The IVIS seamlessly integrates with single- and multi-column ÄKTA Chromatography configurations, enabling fully automated, continuous processes from protein capture chromatography to viral inactivation.

Validation Results: The system demonstrated high precision, achieving pH control within ±0.15 at a target pH of 3 for continuous Protein A eluates. The coiled flow reactor maintained a residence time of 13.5 minutes with a relative width of 0.7, showcasing its capability for reliable and robust performance in continuous downstream processing.

Figure 1. Diagram of the inline viral inactivation system (IVIS) integrated with AKTA Chromatography.

Figure 2. Multi-column chromatograph system (left) where products are eluted into the IVIS system (middle) along with a laptop for adaptive control and data capturing. Viral inactivated products are passed onto further downstream processes (right).

References

1. Lee, J. S. Z., Nguyen, T. D., Zheng, Z. Y., Zhang, W., & Liu, D. (2024). Real‐Time adaptive inline acidification enhances continuous pH control for viral inactivation. Biotechnology Journal, 19(11). https://doi.org/10.1002/biot.202400456
2. O. Yang, M. Qadan, and M. Ierapetritou, “Economic Analysis of Batch and Continuous Biopharmaceutical Antibody Production: A Review,” Journal of Pharmaceutical Innovation 15, no. 1 (2020): 182–200, https://doi.org/10.1007/S12247-018-09370-4/metrics
3. C. L. Burcham, A. J. Florence, and M. D. Johnson (2018). “Continuous Manufacturing in Pharmaceutical Process Development and Manufacturing,” Annual Review of Chemical and Biomolecular Engineering 9 (2018), 253–281, https://doi.org/10.1146/ANNUREV-CHEMBIOENG-060817-084355
4. J. Pollock, J. Coffman, S. V. Ho, and S. S. Farid, “Integrated Continuous Bioprocessing: Economic, Operational, and Environmental Feasibility for Clinical and Commercial AntibodyManufacture,” Biotechnology Progress 33, no. 4 (2017): 854–866, https://doi.org/10.1002/BTPR.2492
5. A. S. Rathore, S. Nikita, G. Thakur, and N. Deore, “Challenges in Process Control for Continuous Processing for Production of Monoclonal Antibody Products,” Current Opinion in Chemical Engineering 31 (2021): 100671, https://doi.org/10.1016/J.COCHE.2021.100671
6. M. S. Hong, A. E. Lu, R. W. Ou, et al., “Model-Based Control for Column-Based Continuous Viral Inactivation of Biopharmaceuticals,” Biotechnology and Bioengineering 118, no. 8 (2021): 3215–3224, https://doi.org/10.1002/BIT.27846
7. G. Thakur, P. Ghumade, and A. S. Rathore, “Process Analytical Technology in Continuous Processing: Model-Based Real Time Control of pH Between Capture Chromatography and Viral Inactivation for Monoclonal Antibody Production,” Journal of Chromatography A 1658 (2021): 462614, https://doi.org/10.1016/J.CHROMA.2021.462614