Transcriptome and proteome analysis of steady‐state in a perfusion CHO cell culture process

Abstract

Long‐term continuous protein production can be reached by perfusion operation. Through the continuous removal of waste metabolites and supply of nutrients, steady‐state (SS) conditions are achieved after a certain transient period, where the conditions inside the reactor are not only uniform in space but also constant in time. Such stable conditions may have beneficial influences on the reduction of product heterogeneities. In this study, we investigated the impact of perfusion cultivation on the intracellular physiological state of a CHO cell line producing a monoclonal antibody (mAb) by global transcriptomics and proteomics. Despite stable viable cell density was maintained right from the beginning of the cultivation time, productivity decrease, and a transition phase for metabolites and product quality was observed before reaching SS conditions. These were traced back to three sources of transient behaviors being hydrodynamic flow rates, intracellular dynamics of gene expression as well as metabolism and cell line instability, superimposing each other. However, 99.4% of all transcripts and proteins reached SS during the first week or were at SS from the beginning. These results demonstrate that the stable extracellular conditions of perfusion lead to SS also of the cellular level.

Insights into the Impact of Rosmarinic Acid on CHO Cell Culture Improvement through Transcriptomics Analysis

“…Similar work reported by Vodopivec et al (2019) compared intracellular metabolomics of different growth phases among three bioreactors scales (10 L, 100 L, and 1000 L) to observe the scale-up effect [20]. As well as the studies in fed-batch cultures, Bertrand et al (2019) applied transcriptomics and proteomics to investigate the influence of long-term perfusion cultivation on the physiological state in an industrial CHO cell line [21]. Overall, omics approaches have contributed to deciphering the complicated CHO cell metabolism better and have aided in the identification of targets for rational genetic engineering and process optimization [22].…”

Proteomic Profiling of IgG1 Producing CHO Cells Using LC/LC-SPS-MS3: The Effects of Bioprocessing Conditions on Productivity and Product Quality

“…Quantitative SPS-MS 3 based LC-MS analysis yielded excellent reproducibility with improved proteome coverage (Bertrand et al, 2019;Lakshmanan et al, 2019) and accurate quantification of proteins (Figure 3) without the need of intense fractionation (Heffner et al, 2017). Statistical data analysis by means of PCA (Figure 4C) indicated that the cellular effects underlying an improved antibody expression at low temperature and low pH differ considerably.…”

Recent Developments in Bioprocessing of Recombinant Proteins: Expression Hosts and Process Development

“…In another study, a single-use fluidic components-based perfusion bioreactor system was developed and enabled the implementation of active environmental control (Bournonville et al, 2019). In a study by Bertrand et al (2019), the impact of perfusion cultivation on the intracellular physiological state of a CHO cell line was investigated, revealing decreased mAb productivity as well as a transition phase for metabolites and product quality before reaching steadystate conditions. For viral vaccine production using anchoragedependent cells (e.g., Vero cells), microcarriers are necessary in the bioreactor.…”