Continuous precipitation‐filtration process for initial capture of a monoclonal antibody product using a four‐stage countercurrent hollow fiber membrane washing step

The significant increase in product titers, coupled with the growing focus on continuous bioprocessing, has renewed interest in using precipitation as a low‐cost alternative to Protein A chromatography for the primary capture of monoclonal antibody (mAb) products. In this work, a commercially releva...

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Bibliographic Details
Published in:	Biotechnology and bioengineering Vol. 121; no. 8; pp. 2258 - 2268
Main Authors:	Minervini, Mirko, Mergy, Matthew, Zhu, Yuncan, Gutierrez Diaz, Mario A., Pointer, Craig, Shinkazh, Oleg, Oppenheim, Sheldon F., Cramer, Steven M., Przybycien, Todd M., Zydney, Andrew L.
Format:	Journal Article
Language:	English
Published:	United States Wiley Subscription Services, Inc 01-08-2024 Wiley
Subjects:	Acetic acid Animals Antibodies, Monoclonal - chemistry Antibodies, Monoclonal - isolation & purification BASIC BIOLOGICAL SCIENCES Bioprocessing Biotechnology & Applied Microbiology Cell culture Cell size Chemical Precipitation CHO Cells Clearances Continuous fibers Countercurrent Distribution - methods Cricetulus Deoxyribonucleic acid Dewatering DNA Filtration - methods Fluid flow Hollow fiber membranes mAb Membrane proteins Microfiltration Monoclonal antibodies monoclonal antibody Optical data processing Particle size distribution precipitation primary capture Protein A Proteins Size distribution tangential flow filtration Washing precipitation monoclonal antibody primary capture tangential flow filtration mAb
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Summary:	The significant increase in product titers, coupled with the growing focus on continuous bioprocessing, has renewed interest in using precipitation as a low‐cost alternative to Protein A chromatography for the primary capture of monoclonal antibody (mAb) products. In this work, a commercially relevant mAb was purified from clarified cell culture fluid using a tubular flow precipitation reactor with dewatering and washing provided by tangential flow microfiltration. The particle morphology was evaluated using an inline high‐resolution optical probe, providing quantitative data on the particle size distribution throughout the precipitation process. Data were obtained in both a lab‐built 2‐stage countercurrent washing system and a commercial countercurrent contacting skid that provided 4 stages of continuous washing. The processes were operated continuously for 2 h with overall mAb yield of 92 ± 3% and DNA removal of nearly 3 logs in the 4‐stage system. The high DNA clearance was achieved by selective redissolution of the mAb using a low pH acetate buffer. Host cell protein clearance was 0.59 ± 0.08 logs, comparable to that based on model predictions. The process mass intensity was slightly better than typical Protein A processes and could be significantly improved by preconcentration of the antibody feed material.
Bibliography:	Disclaimer: This report was prepared as an account of work sponsored by an agency of the United States Government. Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, or favoring by the United States Government or any agency thereof. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States Government or any agency thereof. ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 USDOE Office of Energy Efficiency and Renewable Energy (EERE) National Institute of Standards and Technology (NIST) EE0007613; 70NANB17H002; T32GM141865 National Institutes of Health (NIH)
ISSN:	0006-3592 1097-0290 1097-0290
DOI:	10.1002/bit.28525