Proteomics-based method to comprehensively model the removal of host cell protein impurities.

Autor: Disela R; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands., Keulen D; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands., Fotou E; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands., Neijenhuis T; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands., Le Bussy O; GSK, Technical Research & Development, Rue de l'Institut 89, Rixensart, Belgium., Geldhof G; GSK, Technical Research & Development, Rue de l'Institut 89, Rixensart, Belgium., Pabst M; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands., Ottens M; Department of Biotechnology, Delft University of Technology, Delft, The Netherlands.
Jazyk: angličtina
Zdroj: Biotechnology progress [Biotechnol Prog] 2024 Nov-Dec; Vol. 40 (6), pp. e3494. Date of Electronic Publication: 2024 Jul 17.
DOI: 10.1002/btpr.3494
Abstrakt: Mechanistic models mostly focus on the target protein and some selected process- or product-related impurities. For a better process understanding, however, it is advantageous to describe also reoccurring host cell protein impurities. Within the purification of biopharmaceuticals, the binding of host cell proteins to a chromatographic resin is far from being described comprehensively. For a broader coverage of the binding characteristics, large-scale proteomic data and systems level knowledge on protein interactions are key. However, a method for determining binding parameters of the entire host cell proteome to selected chromatography resins is still lacking. In this work, we have developed a method to determine binding parameters of all detected individual host cell proteins in an Escherichia coli harvest sample from large-scale proteomics experiments. The developed method was demonstrated to model abundant and problematic proteins, which are crucial impurities to be removed. For these 15 proteins covering varying concentration ranges, the model predicts the independently measured retention time during the validation gradient well. Finally, we optimized the anion exchange chromatography capture step in silico using the determined isotherm parameters of the persistent host cell protein contaminants. From these results, strategies can be developed to separate abundant and problematic impurities from the target antigen.
(© 2024 The Author(s). Biotechnology Progress published by Wiley Periodicals LLC on behalf of American Institute of Chemical Engineers.)
Databáze: MEDLINE
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