Multiplex secretome engineering enhances recombinant protein production and purity
| Autor: | Bernhard O. Palsson, Austin W. T. Chiang, Johnny Arnsdorf, Marianne Decker, Sanne Schoffelen, Tune Wulff, Lasse Ebdrup Pedersen, Helene Faustrup Kildegaard, Stefan Kol, Tanja Lyholm Jensen, Jahir M. Gutierrez, Gyun Min Lee, Bjørn G. Voldborg, Anders Holmgaard Hansen, Helen O. Masson, Daniel Ley, Nathan E. Lewis |
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| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine medicine.drug_class Science Cell General Physics and Astronomy CHO Cells Biologics Monoclonal antibody 01 natural sciences Article General Biochemistry Genetics and Molecular Biology Antibodies 03 medical and health sciences Gene Knockout Techniques Cricetulus 010608 biotechnology Monoclonal medicine Protein biosynthesis Computational models Animals Multiplex lcsh:Science Biological Products Chromatography Multidisciplinary biology Chemistry Chinese hamster ovary cell Antibodies Monoclonal High-Throughput Nucleotide Sequencing General Chemistry Recombinant Proteins 030104 developmental biology Secretory protein medicine.anatomical_structure Biochemistry Metabolic Engineering Cell culture biology.protein lcsh:Q Synthetic Biology Antibody Rituximab Biotechnology |
| Zdroj: | Nature communications, vol 11, iss 1 Nature Communications, Vol 11, Iss 1, Pp 1-10 (2020) Kol, S, Ley, D, Wulff, T, Decker, M, Arnsdorf, J, Schoffelen, S, Hansen, A H, Jensen, T L, Gutierrez, J M, Chiang, A W T, Masson, H O, Palsson, B O, Voldborg, B G, Pedersen, L E, Kildegaard, H F, Min Lee, G & Lewis, N E 2020, ' Multiplex secretome engineering enhances recombinant protein production and purity ', Nature Communications, vol. 11, no. 1 . https://doi.org/10.1038/s41467-020-15866-w Nature Communications |
| Popis: | Host cell proteins (HCPs) are process-related impurities generated during biotherapeutic protein production. HCPs can be problematic if they pose a significant metabolic demand, degrade product quality, or contaminate the final product. Here, we present an effort to create a “clean” Chinese hamster ovary (CHO) cell by disrupting multiple genes to eliminate HCPs. Using a model of CHO cell protein secretion, we predict that the elimination of unnecessary HCPs could have a non-negligible impact on protein production. We analyze the HCP content of 6-protein, 11-protein, and 14-protein knockout clones. These cell lines exhibit a substantial reduction in total HCP content (40%-70%). We also observe higher productivity and improved growth characteristics in specific clones. The reduced HCP content facilitates purification of a monoclonal antibody. Thus, substantial improvements can be made in protein titer and purity through large-scale HCP deletion, providing an avenue to increased quality and affordability of high-value biopharmaceuticals. Host cell proteins can contaminate biotherapeutics and compromise and degrade their quality. Here the authors use modelling and CRISPR to delete secreted host proteins in CHO cells, leading to improved monoclonal antibody production and purity. |
| Databáze: | OpenAIRE |
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