Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via "flow‐through affinity chromatography" using peptide‐based adsorbents. Issue 7 (22nd April 2022)

• Record Type: Journal Article
• Authors: Sripada, Sobhana Alekhya; Chu, Wenning; Williams, Taufika Islam; Teten, Matthew A.; Mosley, Brian J.; Carbonell, Ruben G.; Lenhoff, Abraham M.; Cramer, Steven M.; Bill, Jerome; Yigzaw, Yinges; Roush, David J.; Menegatti, Stefano
• Is Part Of: Biotechnology and bioengineering. Volume 119:Issue 7(2022)
• Page Start: 1873
• Page End: 1889
• Publication Date: 2022-04-22
• Subjects: CHO -- flow‐through chromatography -- host cell proteins -- monoclonal antibodies -- peptide‐based adsorbents; Biotechnology -- Periodicals; Bioengineering -- Periodicals; 660.6
• DOI: 10.1002/bit.28096
• Languages: English
• ISSNs: 0006-3592

Abstract

Abstract: The 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. Abstract : A novel purification paradigm based on the selective capture of host cell proteins (HCPs) from CHO cell culture fluids containing monoclonal antibodies (mAbs) was demonstrated using a novel adsorbent (LigaGuard™) developed by Prof. Menegatti and coworkers. Using an ensemble of analytical techniques—including proteomics, enzyme‐linked immunosorbent assay, and analytical chromatography—the authors demonstrated the ability of LigaGuard™ to remove persistent and high‐risk HCPs via "flow‐through affinity chromatography, " thus enabling a next‐generation mAb purification process with superior robustness and performance.