(740h) ‘Omics Profiling of a Continuous Manufacturing CHO Production Platform for Monoclonal Antibodies

Authors: 
Bertrand, V., ETH Zurich
Karst, D., ETH Zurich
Soos, M., University of Chemistry and Technology Prague
Morbidelli, M., ETH Zürich
The manufacturing of biotherapeutics such as recombinant proteins is usually performed in fed-batch mode using Chinese hamster ovary (CHO) cell lines as main expression system. Currently, continuous manufacturing is only preferred if the protein is unstable and prone to chemical or enzymatic degradation. Nevertheless, using perfusion reactors is beneficial for the reduction of quality inconsistencies due to the short residence and steady operation, which is equally favourable for more stable products as monoclonal antibodies (mAbs). Realizing process and product steady-state by systematic process development instead of using empirically driven approaches is one of the key points within the Quality by design (QbD) concept. Thereby, the increase in knowledge about cell physiology is a cornerstone within this development strategy, where â??omics technologies can be helpful to address the regulatory behaviour of the used cell lines.

In this work, transcriptome and proteome profiling of a CHO cell line producing a mAb was performed to investigate the intracellular variability during steady-state of a perfusion reactor. Daily samples were analysed by high-throughput methods depicting the time evolution of several thousand transcripts and proteins. Data mining by different clustering methods allowed the analysis of each individual â??omics study. Furthermore up- and down regulated intracellular processes and pathways responsible for the quality of the secreted mAb were investigated. By the use of bioinformatics databases the biological function was assigned and moreover key functions were characterized by functional annotation. Additionally, to identify differences between the mRNA and protein expression in time, an integrated data analysis of both datasets was completed, as the correlation of the proteomic and transcriptomic expression response is not necessarily given for every regulatory cell process. In conclusion, useful insights into cellular behaviour can be gained by the joint application of â??omics technologies to address the intracellular process consistency of perfusion cell culture.