Transcriptome analysis for the scale-down of a CHO cell fed-batch process

Alsayyari, Abdulaziz A.; Pan, Xiao; Dalm, Ciska; Veen, Jochem W. van der; Vriezen, Nienke; Hageman, Jos A.; Wijffels, René H.; Martens, Dirk E.

doi:10.1016/j.jbiotec.2018.05.012

Cited by 12 publications

(5 citation statements)

References 27 publications

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“…Affymetrix GeneChip CHO Gene 2.1 ST Arrays (Affymetrix, Santa Clara, USA) were used for transcriptome expression profiling. The detailed method was presented in the study of Alsayyari et al In short, the same amount of RNA was labeled by the Whole‐transcript Sense Target Assay (Affymetrix) and hybridized according to the manufacturer's instructions. Quality control and data analysis were done as described in ref.…”

Section: Methodsmentioning

confidence: 99%

“…The metabolic changes associated with the cell size increase were studied in detail using flux balance analysis; however, the cause and molecular mechanisms for the cell size increase remained unclear. Transcriptome analysis has been used as a powerful tool to better understand the physiology of CHO cell lines . The aim of the present study is to obtain more insights into the cause and the molecular mechanisms underlying the CHO cell size increase using transcriptome analysis.…”

Section: Introductionmentioning

confidence: 99%

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Transcriptome Analysis of CHO Cell Size Increase During a Fed-Batch Process

et al. 2018

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In a Chinese Hamster Ovary (CHO) cell fed-batch process, arrest of cell proliferation and an almost threefold increase in cell size occurred, which is associated with an increase in cell-specific productivity. In this study, transcriptome analysis is performed to identify the molecular mechanisms associated with this. Cell cycle analysis reveals that the cells are arrested mainly in the G /G phase. The cell cycle arrest is associated with significant up-regulation of cyclin-dependent kinases inhibitors (CDKNs) and down-regulation of cyclin-dependent kinases (CDKs) and cyclins. During the cell size increase phase, the gene expression of the upstream pathways of mechanistic target of rapamycin (mTOR), which is related to the extracellular growth factor, cytokine, and amino acid conditions, shows a strongly synchronized pattern to promote the mTOR activity. The downstream genes of mTOR also show a synchronized pattern to stimulate protein translation and lipid synthesis. The results demonstrate that cell cycle inhibition and stimulated mTOR activity at the transcriptome level are related to CHO cell size increase. The cell size increase is related to the extracellular nutrient conditions through a number of cascade pathways, indicating that by rational design of media and feeds, CHO cell size can be manipulated during culture processes, which may further improve cell growth and specific productivity.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Transcriptome Analysis of CHO Cell Size Increase During a Fed-Batch Process

et al. 2018

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“…This is conventionally done by keeping a hydrodynamic state constant, e.g. volumetric power input ( P / V L ) ( Klöckner et al, 2012;Catapano et al, 2009 ), mixing time ( Varley and Birch, 1999;Rosseburg et al, 2018 ), impeller tip speed ( Ju and Chase, 1992;Alsayyari et al, 2018 ) or the volumetric mass transfer coefficient k L a ( Xing et al, 2009;Nienow et al, 1996 ). Therefore, it is recommended to hydrodynamically characterize the bioreactors at each scale (recommendation see Meusel et al, 2016 ).…”

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confidence: 99%

Model uncertainty-based evaluation of process strategies during scale-up of biopharmaceutical processes

Möller

Rodríguez

Müller

et al. 2020

Computers & Chemical Engineering

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Model-assisted design of experiments Quality by design a b s t r a c tReliable scale-up of biopharmaceutical production processes is key in Quality by Design. In this study, a model-based workflow is described to evaluate the bioprocess dynamics during process transfer and scale-up computationally. First, a mathematical model describes the bioprocess dynamics of different state variables (e.g., cell density, titer). Second, the model parameter probability distributions are determined at different scales due to measurement uncertainty. Third, the quantified parameter distributions are statistically compared to evaluate if the process dynamics have been changed. This workflow was tested for the scale-up of an antibody-producing CHO fed-batch process. Significant differences were identified between the process development (30 ml) and implementation (250 ml) scale, and the feeding strategy was validated using model-assisted Design of Experiments. Then, the validated process strategy was successfully scaled up to 2 l laboratory and 50 l pilot scale. In summary, the proposed workflow enables a knowledge-driven evaluation tool for bioprocess development.

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“…FB) and a high-inoculation fed-batch using triplicate cultivations for all approaches. Possible scalability-relevant factors have been evaluated in other works [21]. Basic parameters for the bioreactors were: temperature of 36.8 • C; pH of 7.1, adjusted by CO 2 sparging; and DO of 40% by additional O 2 gassing and agitation at 1300 rpm.…”

Section: Small-scale Cultivationmentioning

confidence: 99%

Triple Space-Time Yield in Discontinuous Antibody Biomanufacturing by Combination of Synergetic Process Intensification Strategies

Reger,

Saballus,

Kampmann

et al. 2023

Bioengineering

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Monoclonal antibodies are the workhorse of the pharmaceutical industry due to their potential to treat a variety of different diseases while providing high specificity and efficiency. As a consequence, a variety of production processes have been established within the biomanufacturing industry. However, the rapidly increasing demand for therapeutic molecules amid the recent COVID-19 pandemic demonstrated that there still is a clear need to establish novel, highly productive, and flexible production processes. Within this work, we designed a novel discontinuous process by combining two intensification strategies, thus increasing inoculation density and media exchange via a fluidized bed centrifuge, to fulfill the need for a flexible and highly productive production process for therapeutic molecules. To establish this new process, firstly, a small-scale experiment was conducted to verify synergies between both intensification strategies, followed by a process transfer towards the proof-of-concept scale. The combination of these two-process intensification measures revealed overall synergies resulting in decreased process duration (−37%) and strongly enhanced product formation (+116%) in comparison to the not-intensified standard operation. This led to an impressive threefold increase in space-time yield, while only negligible differences in product quality could be observed. Overall, this novel process not only increases the ways to react to emergency situations thanks to its flexibility and possible short development times, but also represents a possible alternative to the current established processes due to high increases in productivity, in comparison to standard fed-batch operations.

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Transcriptome analysis for the scale-down of a CHO cell fed-batch process

Cited by 12 publications

References 27 publications

Transcriptome Analysis of CHO Cell Size Increase During a Fed-Batch Process

Transcriptome Analysis of CHO Cell Size Increase During a Fed-Batch Process

Model uncertainty-based evaluation of process strategies during scale-up of biopharmaceutical processes

Triple Space-Time Yield in Discontinuous Antibody Biomanufacturing by Combination of Synergetic Process Intensification Strategies

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