Multiple steady states with distinct cellular metabolism in continuous culture of mammalian cells

Europa, Anna F.; Gambhir, Anshu; Fu, Peng; Hu, Wei Shou

doi:10.1002/(sici)1097-0290(20000105)67:1<25::aid-bit4>3.0.co;2-k

Cited by 165 publications

(92 citation statements)

References 14 publications

(16 reference statements)

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“…Controlling glucose at low levels has been demonstrated to be an effective approach in controlling lactate level in mammalian cell cultures [28,[39][40][41]. Similar to the approach of supplementing glucose with other carbon sources, controlling glucose at low levels limits the availability of glucose for the rapid glycolysis pathway and forces lactate into the TCA cycle for fully oxidative metabolism [28,40].…”

Section: Impact Of Control Strategy On Lactate Metabolismmentioning

confidence: 99%

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Hoshan

Chen

2016

Bioprocess Biosyst Eng

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In this study, we discussed the development and optimization of an intensified CHO culture process, highlighting medium and control strategies to improve lactate metabolism. A few strategies, including supplementing glucose with other sugars (fructose, maltose, and galactose), controlling glucose level at <0.2 mM, and supplementing medium with copper sulfate, were found to be effective in reducing lactate accumulation. Among them, copper sulfate supplementation was found to be critical for process optimization when glucose was in excess. When copper sulfate was supplemented in the new process, two-fold increase in cell density (66.5 ± 8.4 × 10(6) cells/mL) and titer (11.9 ± 0.6 g/L) was achieved. Productivity and product quality attributes differences between batch, fed-batch, and concentrated fed-batch cultures were discussed. The importance of process and cell metabolism understanding when adapting the existing process to a new operational mode was demonstrated in the study.

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Section: Impact Of Control Strategy On Lactate Metabolismmentioning

confidence: 99%

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Hoshan

Chen

2016

Bioprocess Biosyst Eng

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“…One key advantage of stoichiometric models is that they can account for competing reactions, which is useful for studying the relative activity of certain pathways under various culture conditions and this has been their major use. For example, the existence of more than one physiological steady state, known as steady-state multiplicity, has been observed by a number of investigators using stoichiometric models and metabolic flux analysis (Follstad et al 1999;Europa et al 2000;Cruz et al 1999;Zhou et al 1997;Linz et al 1997;Paredes et al 1999).…”

Section: Single Cell Modelsmentioning

confidence: 99%

Modelling of Mammalian Cells and Cell Culture Processes

Sidoli¹,

Mantalaris

Asprey³

2004

Cytotechnology

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Mammalian cell cultures represent the major source for a number of very high-value biopharmaceutical products, including monoclonal antibodies (MAbs), viral vaccines, and hormones. These products are produced in relatively small quantities due to the highly specialised culture conditions and their susceptibility to either reduced productivity or cell death as a result of slight deviations in the culture conditions. The use of mathematical relationships to characterise distinct parts of the physiological behaviour of mammalian cells and the systematic integration of this information into a coherent, predictive model, which can be used for simulation, optimisation, and control purposes would contribute to efforts to increase productivity and control product quality. Models can also aid in the understanding and elucidation of underlying mechanisms and highlight the lack of accuracy or descriptive ability in parts of the model where experimental and simulated data cannot be reconciled. This paper reviews developments in the modelling of mammalian cell cultures in the last decade and proposes a future direction -the incorporation of genomic, proteomic, and metabolomic data, taking advantage of recent developments in these disciplines and thus improving model fidelity. Furthermore, with mammalian cell technology dependent on experiments for information, model-based experiment design is formally introduced, which when applied can result in the acquisition of more informative data from fewer experiments. This represents only part of a broader framework for model building and validation, which consists of three distinct stages: theoretical model assessment, model discrimination, and model precision, which provides a systematic strategy from assessing the identifiability and distinguishability of a set of competing models to improving the parameter precision of a final validated model. NomenclatureC ij -component i concentration in jth model compartment; R ijk -transport rate of component i into or out of compartment j from the kth source or sink compartment; N in -number of source compartments; N outnumber of sink compartments; r ijl -reaction rate of component i in compartment j in the lth i-generating or i-consuming reaction; N gen -number of reactions generating component i; N con -number of reactions consuming component i; -specific growth rate; app max -apparent maximum specific growth rate; K dspecific death rate; N v -viable cell number; N t -total cell number; D -dilution rate; -parameter; 0 -parameter; -parameter; m -cell mass; m 0 -mass of a dividing cell; t -time; N -cell number; r -single cell growth rate; S -nutrient concentration; s -nutrient concentration vector; À -cell transition rate; ppartition probability density function; Y -yield coefficient; f (m) -the division probability density function; B -coefficient for the beta distribution incorporating the gamma function; x max -vector of maximum XPS 120742 (CYTO) -ms-code CYTO853 -product element 5254543 -23 September 2004 -Newgen

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“…To better control cell metabolism and avoid using of animal-derived raw materials and ill-defined hydrolysates, much progress has been made in understanding cell nutrient requirements and development of chemically defined medium including feed formulations over the last 10 years (Wurm 2004;Franek et al 2000;Huang et al 2010;Young 2013). More recently, metabolic flux analysis (MFA) has become a useful tool in medium development for hybridoma Europa et al 2000;Gambhir et al 2003) and CHO cell cultures (Nyberg et al 1999;Altamirano et al 2006), which can help quantifying nutrient demands and metabolite production in cell culture (Zhao et al 2009). …”

Section: Introductionmentioning

confidence: 99%

Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

Zhang

Wang

et al. 2016

Bioresour. Bioprocess.

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Background: Optimization of chemically defined medium has been a critical way to produce monoclonal antibody. Usually, amount of glutamine was added into the feed medium, but a half of asparagine was added. Our study found that asparagine was important in the antibody production phase. Increasing the ratio of asparagine to glutamine in feed medium for enhancement of antibody production in CHO-DHFR cell culture would be an efficient way. Results:We optimized the total amount and the ratio of the two vital amino acids in feed medium to increase antibody production. In this work, we have demonstrated that feeding medium of high ratio between asparagine and glutamine (FB-H) can enhance the cell density after reaching the stationary phase. Moreover, FB-H was shown to improve cell maintenance, and increased the antibody production. The metabolic flux analysis proved that ratio of asparagine to glutamine had little influence on glycolysis. Furthermore, the TCA cycle of FB-H was enhanced by 20 % compared to that of low ratio of asparagine to glutamine (FB-L). And the energy metabolism of FB-H was 22.6 % higher than that of FB-L. For the later, lactate can be less produced in FB-H. Conclusions:We should improve the ratio between asparagine and glutamine in feed medium properly under the premise of no influence on cell growth to achieve high mAb producing goal.

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Multiple steady states with distinct cellular metabolism in continuous culture of mammalian cells

Cited by 165 publications

References 14 publications

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations

Modelling of Mammalian Cells and Cell Culture Processes

Responses of CHO-DHFR cells to ratio of asparagine to glutamine in feed media: cell growth, antibody production, metabolic waste, glutamate, and energy metabolism

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