Proteomics analysis of altered cellular metabolism induced by insufficient copper level

Kang, Sohye; Xiao, Gang; Ren, Da; Zhang, Zhongqi; Le, Nicole; Trentalange, Michael; Gupta, Shivani; Lin, Henry; Bondarenko, Pavel V.

doi:10.1016/j.jbiotec.2014.08.001

Cited by 11 publications

(15 citation statements)

References 54 publications

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“…In previous studies, researchers found that increased copper led to high productivity of the CHO cell process with a decrease (consumption) of lactate in the stationary region. It was reasoned that the higher copper level affected copper binding to the COX proteins, thus reducing ROS produced in the mitochondria . In the present study, we did not find lactate consumption for the 5‐kL scale.…”

Section: Discussioncontrasting

confidence: 64%

“…For the 20‐L scale, the lactate concentration decreased after 120 hours; however, again, there was no difference between the copper concentration levels. Addition of copper has previously been shown to result in consumption of lactate, which is generally considered a desirable phenotype for improvement of production and viability . Our results are at variance with those reported in the literature; however, it was pointed in a recent paper that the lactate behavior with added copper is dependent on the cell line and conditions used .…”

Section: Resultscontrasting

confidence: 55%

“…Copper, a well‐known trace metal of cell culture media, serves as a cofactor of many enzymes controlling their functional states and activity levels . There have been several previous reports describing the effects of copper levels on the productivity of CHO cells, albeit for low volume reactors or shake flasks .…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Combined metabolomics and proteomics reveals hypoxia as a cause of lower productivity on scale‐up to a 5000‐liter CHO bioprocess

Gao

Ray

Dai

et al. 2016

Biotechnology Journal

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Large-scale bioprocessing is key to the successful manufacturing of a biopharmaceutical. However, cell viability and productivity are often lower in the scale-up from laboratory to production. In this study, we analyzed CHO cells, which showed lower percent viabilities and productivity in a 5-KL production scale bioreactor compared to a 20-L bench-top scale under seemingly identical process parameters. An increase in copper concentration in the media from 0.02 µM to 0.4 µM led to a doubling of percent viability in the production scale albeit still at a lower level than the bench-top scale. Combined metabolomics and proteomics revealed the increased copper reduced the presence of reactive oxygen species (ROS) in the 5-KL scale process. The reduction in oxidative stress was supported by the increased level of glutathione peroxidase in the lower copper level condition. The excess ROS was shown to be due to hypoxia (intermittent), as evidenced by the reduction in fibronectin with increased copper. The 20-L scale showed much less hypoxia and thus less excess ROS generation, resulting in little to no impact to productivity with the increased copper in the media. The study illustrates the power of 'Omics in aiding in the understanding of biological processes in biopharmaceutical production.

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Section: Discussioncontrasting

confidence: 64%

Section: Resultscontrasting

confidence: 55%

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Combined metabolomics and proteomics reveals hypoxia as a cause of lower productivity on scale‐up to a 5000‐liter CHO bioprocess

Gao

Ray

Dai

et al. 2016

Biotechnology Journal

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show abstract

“…One particular trace element, copper, has been identified to be necessary in promoting cell growth and inducing lactate consumption in a wide range of CHO cell lines [20][21][22][23]. Cultures with copper deficiency have been found to lose the ability in consuming lactate in the stationary phase.…”

Section: Impact Of Medium Components On Lactate Metabolismmentioning

confidence: 99%

“…Copper is an important cofactor for cytochrome c oxidase, the terminal enzyme in the mitochondrial electron transport chain. Decrease in the protein levels of cytochrome c oxidase subunits has been observed under the copper-deficient condition [22]. Studies also suggested that low mitochondrial oxidative capacity will lead to high lactate production [25], and lactate-producing cultures could have impaired mitochondrial and energy metabolism due to insufficient copper level [21].…”

Section: Impact Of Medium Components 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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Mechanisms driving the lactate switch in Chinese hamster ovary cells

Hartley

Walker

Chung

et al. 2018

Biotech & Bioengineering

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The metabolism of Chinese Hamster Ovary (CHO) cells in a production environment has been extensively investigated. However, a key metabolic transition, the switch from lactate production to lactate consumption, remains enigmatic. Though commonly observed in CHO cultures, the mechanism(s) by which this metabolic shift is triggered is unknown. Despite this, efforts to control the switch have emerged due to the association of lactate consumption with improved cell growth and productivity. This review aims to consolidate current theories surrounding the lactate switch. The influence of pH, NAD /NADH, pyruvate availability and mitochondrial function on lactate consumption are explored. A hypothesis based on the cellular redox state is put forward to explain the onset of lactate consumption. Various techniques implemented to control the lactate switch, including manipulation of the culture environment, genetic engineering, and cell line selection are also discussed.

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Proteomics analysis of altered cellular metabolism induced by insufficient copper level

Cited by 11 publications

References 54 publications

Combined metabolomics and proteomics reveals hypoxia as a cause of lower productivity on scale‐up to a 5000‐liter CHO bioprocess

Combined metabolomics and proteomics reveals hypoxia as a cause of lower productivity on scale‐up to a 5000‐liter CHO bioprocess

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

Mechanisms driving the lactate switch in Chinese hamster ovary cells

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