Characterization of dynamic regulation in Chinese hamster ovary (CHO) cell cultures in the late exponential phase

Sha, Sha; Kuang, Bingyu; Yoon, Seongkyu

doi:10.1016/j.bej.2020.107897

Cited by 7 publications

(5 citation statements)

References 49 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast, the exchange of media in the stationary phase triggers a different cellular behavior with prolonged viability and without proliferation since the majority of cells have already transitioned into the non-dividing state. The transition from the exponential to the stationary phase is not fully analyzed up to this time point but is associated with the accumulation of inhibiting molecules in the supernatant, resulting in several intracellular changes starting at the transcriptional level ( Hernández Bort et al, 2012 ; Sha et al, 2021 ), thereby likely impacting the metabolic flux in different pathways, for example, pentose phosphate pathway and TCA cycle ( Sengupta et al, 2011 ; Templeton et al, 2013 ), which is also shown by a change in the holistic cellular behavior like cell size increase ( Pan et al, 2017 ). The overall changes in the cell culture at transition into the stationary phase were permanent for our conducted studies, even with supply of fresh media.…”

Section: Discussionmentioning

confidence: 99%

Boosting Productivity for Advanced Biomanufacturing by Re-Using Viable Cells

Reger

Saballus

Matuszczyk

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Monoclonal antibodies (mAb) have gained enormous therapeutic application during the last decade as highly efficient and flexible tools for the treatment of various diseases. Despite this success, there remain opportunities to drive down the manufacturing costs of antibody-based therapies through cost efficiency measures. To reduce production costs, novel process intensification methods based on state-of-the-art fed-batch and perfusion have been implemented during the last few years. Building on process intensification, we demonstrate the feasibility and benefits of a novel, innovative hybrid process that combines the robustness of a fed-batch operation with the benefits of a complete media exchange enabled through a fluidized bed centrifuge (FBC). In an initial small-scale FBC-mimic screening, we investigated multiple process parameters, resulting in increased cell proliferation and an elongated viability profile. Consecutively, the most productive process scenario was transferred to the 5-L scale, further optimized and compared to a standard fed-batch process. Our data show that the novel hybrid process enables significantly higher peak cell densities (163%) and an impressive increase in mAb amount of approximately 254% while utilizing the same reactor size and process duration of the standard fed-batch operation. Furthermore, our data show comparable critical quality attributes (CQAs) between the processes and reveal scale-up possibilities and no need for extensive additional process monitoring. Therefore, this novel process intensification strategy yields strong potential for transfer into future industrial manufacturing processes.

show abstract

Section: Discussionmentioning

confidence: 99%

Boosting Productivity for Advanced Biomanufacturing by Re-Using Viable Cells

Reger

Saballus

Matuszczyk

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…It is important to note that the threshold is user-defined. The fold-changes used in previous studies varied from 1.2 to 1.5 or higher [27][28][29][30][31]. A web server KOBAS was used to perform the pathway enrichment analysis using a set of identified DEGs [32].…”

Section: Metabolic Pathway Analysismentioning

confidence: 99%

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

Huang

Tian

et al. 2022

Processes

View full text Add to dashboard Cite

The use of antioxidants in Chinese hamster ovary (CHO) cell cultures to improve monoclonal antibody production has been a topic of great interest. Nevertheless, the antioxidants do not have consistent benefits of production improvement, which might be cell line specific and/or process specific. In this work, we investigated how treatment with the antioxidant rosmarinic acid (RA) improved cell growth and titer in CHO cell cultures using transcriptomics. In particular, transcriptomics analysis indicated that RA treatment modified gene expression and strongly affected the MAPK and PI3K/Akt signaling pathways, which regulate cell survival and cell death. Moreover, it was observed that these signaling pathways, which had been identified to be up-regulated on day 2 and day 6 by RA, were also up-regulated over time (from initial growth phase day 2 to slow growth or protein production phase day 6) in both conditions. In summary, this transcriptomics analysis provides insights into the role of the antioxidant RA in industrial cell culture processes. The current study also represents an example in the industry of how omics can be applied to gain an in-depth understanding of CHO cell biology and to identify critical pathways that can contribute to cell culture process improvement and cell line engineering.

show abstract

“…The explanation for lack of continuous process characteristics, i.e., the decreasing viabilities and cell proliferation, within these cultures could be an irreversible transition of the cells into the stationary phase without further proliferation potential. This transition can be triggered by the accumulation of metabolic byproducts, which ultimately limit further proliferation and finally reduce viabilities ( Lao and Toth, 1997 ; Mulukutla et al, 2017 ; Sha et al, 2021 ). Another explanation for this irreversible transition of the respective cultures (30 d2; 20 d3) could be a limitation of nutrients ( Xie and Wang, 1994 ; Xie et al, 1997 ; Lu et al, 2013 ).…”

Section: Discussionmentioning

confidence: 99%

A novel hybrid bioprocess strategy addressing key challenges of advanced biomanufacturing

Reger,

Saballus,

Kappes

et al. 2023

Front. Bioeng. Biotechnol.

View full text Add to dashboard Cite

Monoclonal antibodies (mAb) are commonly manufactured by either discontinuous operations like fed-batch (FB) or continuous processes such as steady-state perfusion. Both process types comprise opposing advantages and disadvantages in areas such as plant utilization, feasible cell densities, media consumption and process monitoring effort. In this study, we show feasibility of a promising novel hybrid process strategy that combines beneficial attributes of both process formats. In detail, our strategy comprises a short duration FB, followed by a fast media exchange and cell density readjustment, marking the start of the next FB cycle. Utilizing a small-scale screening tool, we were able to identify beneficial process parameters, including FB interval duration and reinoculation cell density, that allow for multiple cycles of the outlined process in a reproducible manner. In addition, we could demonstrate scalability of the process to a 5L benchtop system, using a fluidized-bed centrifuge as scalable media exchange system. The novel process showed increased productivity (+217%) as well as longer cultivation duration, in comparison to a standard FB with a significantly lower media consumption per produced product (−50%) and a decreased need for process monitoring, in comparison to a perfusion cultivation. Further, the process revealed constant glycosylation pattern in comparison to the perfusion cultivation and has strong potential for further scale-up, due to the use of fully scalable cultivation and media exchange platforms. In summary, we have developed a novel hybrid process strategy that tackles the key challenges of current biomanufacturing of either low productivity or high media consumption, representing a new and innovative approach for future process intensification efforts.

show abstract

Characterization of dynamic regulation in Chinese hamster ovary (CHO) cell cultures in the late exponential phase

Cited by 7 publications

References 49 publications

Boosting Productivity for Advanced Biomanufacturing by Re-Using Viable Cells

Boosting Productivity for Advanced Biomanufacturing by Re-Using Viable Cells

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

A novel hybrid bioprocess strategy addressing key challenges of advanced biomanufacturing

Contact Info

Product

Resources

About