The publishing of the CHO genome in 2013 was the first step towards advancing our knowledge of CHO cell biology. With a deeper knowledge of CHO cell biology, cellular engineering strategies can be developed to target pathways and proteins that are associated with phenotypes of interest. Little progress has been made towards understanding the intracellular pathways that contribute to creating industrially desirable phenotypes in CHO cells.
Ivcd cell culture free#
Optimising growth, titre and specific productivity of these cells has long been an area of interest among the pharmaceutical industry however, the vast majority of improvements to date can be attributed to optimised feeding strategies and adaption to serum free medium.
This investigation highlights key pathways for targeted engineering to generate desirable CHO cell phenotypes for biotherapeutic production.ĬHO cells are the most frequently used host cell line for the production of therapeutic proteins due to their ability to produce human like post-translational modifications, their high level of approval among regulatory authorities and their stable transgene expression. ER to Golgi vesicle mediated transport was found to be highly expressed in extended culture VCD CDCLs while networks involving endocytosis and oxidative stress response were significantly downregulated. We identified key pathways in DNA replication, mitotic cell cycle and evasion of p53 mediated apoptosis in high peak VCD clonally derived cell lines (CDCLs). In this study we implemented label-free LC-MS/MS proteomic profiling of IgG4 producing CHO cell lines throughout the duration of the cell culture to identify differentially expressed (DE) proteins and intracellular pathways associated with the high peak viable cell density (VCD) and extended culture VCD phenotypes. The ability to achieve high peak viable cell density earlier in CHO cell culture and maintain an extended cell viability throughout the production process is highly desirable to increase recombinant protein yields, reduce host cell impurities for downstream processing and reduce the cost of goods.