2022 article
Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via "flow-through affinity chromatography" using peptide-based adsorbents
Sripada, S. A., Chu, W., Williams, T. I., Teten, M. A., Mosley, B. J., Carbonell, R. G., … Menegatti, S. (2022, April 22). BIOTECHNOLOGY AND BIOENGINEERING.
author keywords: CHO; flow-through chromatography; host cell proteins; monoclonal antibodies; peptide-based adsorbents
MeSH headings : Animals; Antibodies, Monoclonal / chemistry; CHO Cells; Cell Culture Techniques; Chromatography, Affinity / methods; Cricetinae; Cricetulus; Peptides / chemistry; Proteomics / methods
TL;DR:
An ensemble of peptide ligands that target the HCPs in Chinese hamster ovary (CHO) cell culture fluids and enable mAb purification via flow‐through affinity chromatography are introduced and their integration into LigaGuard™, an affinity adsorbent featuring an equilibrium binding capacity.
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UN Sustainable Development Goal Categories
3. Good Health and Well-being
(Web of Science)
Source: Web Of Science
Added: May 10, 2022
AbstractThe growth of advanced analytics in manufacturing monoclonal antibodies (mAbs) has highlighted the challenges associated with the clearance of host cell proteins (HCPs). Of special concern is the removal of “persistent” HCPs, including immunogenic and mAb‐degrading proteins, that co‐elute from the Protein A resin and can escape the polishing steps. Responding to this challenge, we introduced an ensemble of peptide ligands that target the HCPs in Chinese hamster ovary (CHO) cell culture fluids and enable mAb purification via flow‐through affinity chromatography. This study describes their integration into LigaGuard™, an affinity adsorbent featuring an equilibrium binding capacity of ~30 mg of HCPs per mL of resin as well as dynamic capacities up to 16 and 22 mg/ml at 1‐ and 2‐min residence times, respectively. When evaluated against cell culture harvests with different mAb and HCP titers and properties, LigaGuard™ afforded high HCP clearance, with logarithmic removal values (LRVs) up to 1.5, and mAb yield above 90%. Proteomic analysis of the effluents confirmed the removal of high‐risk HCPs, including cathepsins, histones, glutathione‐S transferase, and lipoprotein lipases. Finally, combining LigaGuard™ for HCP removal with affinity adsorbents for product capture afforded a global mAb yield of 85%, and HCP and DNA LRVs > 4.