Multiplex secretome engineering enhances recombinant protein production and purity

• Published in: Nature communications Vol. 11; no. 1; p. 1908
• Main Authors: Kol, Stefan, Ley, Daniel, Wulff, Tune, Decker, Marianne, Arnsdorf, Johnny, Schoffelen, Sanne, Hansen, Anders Holmgaard, Jensen, Tanja Lyholm, Gutierrez, Jahir M., Chiang, Austin W. T., Masson, Helen O., Palsson, Bernhard O., Voldborg, Bjørn G., Pedersen, Lasse Ebdrup, Kildegaard, Helene Faustrup, Lee, Gyun Min, Lewis, Nathan E.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 20.04.2020; Nature Publishing Group; Nature Portfolio
• Subjects: 13/31; 38/91; 45; 631/114/2397; 631/61/318; 631/61/338/552; 631/61/51; 82/1; 82/16; 82/80; 82/83; Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - isolation & purification; Biological Products; Cell lines; CHO Cells; Chromatography; Cricetulus; CRISPR; Gene Knockout Techniques; High-Throughput Nucleotide Sequencing; Humanities and Social Sciences; Impurities; Metabolic Engineering - methods; Monoclonal antibodies; multidisciplinary; Protein engineering; Proteins; Purity; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Rituximab; Science; Science (multidisciplinary); Secretome; Synthetic Biology
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-020-15866-w

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.