Multiplex secretome engineering enhances recombinant protein production and purity

Kol, S.; Ley, D.; Wulff, T.; Decker, M.; Arnsdorf, J.; Gutierrez, J. M.; Chiang, A. W.; Pedersen, L. E.; Kildegaard, H. F.; Lee, G. M.; Lewis, N.

doi:10.1101/647214

bioRxiv · 10.1101/647214

Multiplex secretome engineering enhances recombinant protein production and purity

Kol, S.·Ley, D.·Wulff, T.·Decker, M.·Arnsdorf, J.·Gutierrez, J. M.·Chiang, A. W.·Pedersen, L. E.·Kildegaard, H. F.·Lee, G. M.·Lewis, N.

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Abstract

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 predicted the elimination of unnecessary HCPs could have a non-negligible impact on protein production. We analyzed the total HCP content of 6-protein, 11-protein, and 14-protein knockout clones and characterized their growth in shake flasks and bioreactors. These cell lines exhibited a substantial reduction in total HCP content (40%-70%). We also observed higher productivity and improved growth characteristics, in specific clones. With the reduced HCP content, protein A and ion exchange chromatography more efficiently purified a monoclonal antibody (mAb) produced in these cells during a three-step purification process. 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.

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Kol, S., Ley, D., Wulff, T., Decker, M., Arnsdorf, J., Gutierrez, J. M., Chiang, A. W., Pedersen, L. E., Kildegaard, H. F., Lee, G. M., Lewis, N.. 2019-05-24. Multiplex secretome engineering enhances recombinant protein production and purity. https://doi.org/10.1101/647214

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