Multiplex secretome engineering enhances recombinant protein production and purity

Nat Commun. 2020 Apr 20;11(1):1908. doi: 10.1038/s41467-020-15866-w.

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 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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / isolation & purification
  • Biological Products
  • CHO Cells
  • Chromatography
  • Cricetulus
  • Gene Knockout Techniques
  • High-Throughput Nucleotide Sequencing
  • Metabolic Engineering / methods*
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Rituximab
  • Synthetic Biology

Substances

  • Antibodies, Monoclonal
  • Biological Products
  • Recombinant Proteins
  • Rituximab