Computational approach for understanding and improving GS-NS0 antibody production under hyperosmotic conditions

Ho, Ying Swan; Kiparissides, Alexandros; Pistikopoulos, Efstratios N.; Mantalaris, Athanasios

doi:10.1016/j.jbiosc.2011.08.022

Cited by 15 publications

(17 citation statements)

References 39 publications

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“…Although osmolarity has not been measured, arguably the 40% more concentrated feed will present an increased extracellular space osmolarity than F_all. This would be in agreement with previous studies showing that increased osmolarity correlates with higher specific productivity (Ho et al, ; Kim et al, ; Takagi et al, ).…”

Section: Resultssupporting

confidence: 94%

A framework for the systematic design of fed‐batch strategies in mammalian cell culture

Kyriakopoulos

Kontoravdi

2014

Biotech & Bioengineering

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A methodology to calculate the required amount of amino acids (a.a.) and glucose in feeds for animal cell culture from monitoring their levels in batch experiments is presented herein. Experiments with the designed feeds on an antibody-producing Chinese hamster ovary cell line resulted in a 3-fold increase in titer compared to batch culture. Adding 40% more nutrients to the same feed further increases the yield to 3.5 higher than in batch culture. Our results show that above a certain threshold there is no linear correlation between nutrient addition and the integral of viable cell concentration. In addition, although high ammonia levels hinder cell growth, they do not appear to affect specific antibody productivity, while we hypothesize that high extracellular lactate concentration is the cause for the metabolic shift towards lactate consumption for the cell line used. Overall, the performance of the designed feeds is comparable to that of a commercial feed that was tested in parallel. Expanding this approach to more nutrients, as well as changing the ratio of certain amino acids as informed by flux balance analysis, could achieve even higher yields.

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Section: Resultssupporting

confidence: 94%

A framework for the systematic design of fed‐batch strategies in mammalian cell culture

Kyriakopoulos

Kontoravdi

2014

Biotech & Bioengineering

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“…However, the presented model can aid in the selection of optimal proliferation control strategies accounting for culture heterogeneity, which is linked to cell growth limitations, cell cycle distribution, and cell cycle-specific productivity. A key aspect of the presented model is its ability to accommodate for various proliferation control strategies, such as temperature shifts (temperature dependent cyclin/DNA growth rates), variation of medium osmolarity (slowing growth and increasing productivity) [78, 79], and chemical approaches for cell cycle arrest (shown herein). The model could be used to optimise different targets, including the time for arrest induction (either by temperature shifts or chemical addition), maximization of the integral of viable cells (temperature, osmotic effects), or maximizing productivity (temperature shifts, osmotic effects, or chemical addition).…”

Section: Discussionmentioning

confidence: 99%

Cyclin and DNA Distributed Cell Cycle Model for GS-NS0 Cells

et al. 2015

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Mammalian cell cultures are intrinsically heterogeneous at different scales (molecular to bioreactor). The cell cycle is at the centre of capturing heterogeneity since it plays a critical role in the growth, death, and productivity of mammalian cell cultures. Current cell cycle models use biological variables (mass/volume/age) that are non-mechanistic, and difficult to experimentally determine, to describe cell cycle transition and capture culture heterogeneity. To address this problem, cyclins—key molecules that regulate cell cycle transition—have been utilized. Herein, a novel integrated experimental-modelling platform is presented whereby experimental quantification of key cell cycle metrics (cell cycle timings, cell cycle fractions, and cyclin expression determined by flow cytometry) is used to develop a cyclin and DNA distributed model for the industrially relevant cell line, GS-NS0. Cyclins/DNA synthesis rates were linked to stimulatory/inhibitory factors in the culture medium, which ultimately affect cell growth. Cell antibody productivity was characterized using cell cycle-specific production rates. The solution method delivered fast computational time that renders the model’s use suitable for model-based applications. Model structure was studied by global sensitivity analysis (GSA), which identified parameters with a significant effect on the model output, followed by re-estimation of its significant parameters from a control set of batch experiments. A good model fit to the experimental data, both at the cell cycle and viable cell density levels, was observed. The cell population heterogeneity of disturbed (after cell arrest) and undisturbed cell growth was captured proving the versatility of the modelling approach. Cell cycle models able to capture population heterogeneity facilitate in depth understanding of these complex systems and enable systematic formulation of culture strategies to improve growth and productivity. It is envisaged that this modelling approach will pave the model-based development of industrial cell lines and clinical studies.

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“…The heavy and light chain assembly of the mAb from transcription to translation was simulated using mass action kinetics (see Terminology) based on a previously suggested model structure . In a follow‐up study, the same group used the model to predict the optimal hyperosmotic induction time for increasing specific antibody productivity . Although the model accurately predicted mAb concentration under different induction times used in calibration experiments, it did not perform well at higher or lower osmotic conditions .…”

Section: Mammalian Cell Culture Kinetic Modelsmentioning

confidence: 99%

“…To date, most of the published kinetic models only encompass culture behavior via intracellular processes at various levels of detail, mainly due to the needs of designing relevant media and feeding strategies, and/or identifying suitable targets for host cell engineering to control the product yield, quality, and cellular growth rates. Comparatively, only a handful of models incorporated other cell culture conditions such as temperature, pH, dissolved oxygen (DO), and osmolality . While temperature, pH, and dissolved oxygen are monitored and controlled during the culture, they can vary within an acceptable range considering the spatial heterogeneity in large‐scale industrial settings which contribute to greater variability in the cell culture environment.…”

Section: Future Directions Of Mammalian Kinetic Modelsmentioning

confidence: 99%

Kinetic Modeling of Mammalian Cell Culture Bioprocessing: The Quest to Advance Biomanufacturing

Kyriakopoulos

Ang

Huang

et al. 2017

Biotechnology Journal

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Kinetic modeling is the most suitable framework to describe the dynamic behavior of mammalian cell culture although its industrial application is still in its infancy. Herein, the authors reviewed mammalian bioprocess relevant kinetic models, and found that the simple unstructured-unsegregated approach utilizing empirical Monod-type kinetics based on limiting substrates and inhibitory metabolites is commonly used due to its traceability and simple formalism. Notably, the available kinetic models are typically small to moderate in size, and the development of large-scale models is severely hampered by the scarcity of kinetic data and limitations in current parameter estimation methods. The recent availability of abundant high-throughput multi-omics datasets from mammalian cell cultures have now paved the way to improve parameterization of kinetic models, and integrate regulatory, signaling, and product quality related intracellular events, as well as cellular metabolism within the modeling framework. Ultimately, the authors foresee that multi-scale modeling is the way forward in building predictive kinetic models of mammalian cell culture to advance biomanufacturing.

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Computational approach for understanding and improving GS-NS0 antibody production under hyperosmotic conditions

Cited by 15 publications

References 39 publications

A framework for the systematic design of fed‐batch strategies in mammalian cell culture

A framework for the systematic design of fed‐batch strategies in mammalian cell culture

Cyclin and DNA Distributed Cell Cycle Model for GS-NS0 Cells

Kinetic Modeling of Mammalian Cell Culture Bioprocessing: The Quest to Advance Biomanufacturing

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