Metabolomics-based identification of apoptosis-inducing metabolites in recombinant fed-batch CHO culture media

Chong, William P. K.; Yusufi, Faraaz Noor Khan; Lee, Dong-Yup; Reddy, Satty G.; Wong, Niki S.C.; Heng, Chew-Kiat; Yap, Miranda G.S.; Ho, Ying Swan

doi:10.1016/j.jbiotec.2010.12.010

Cited by 55 publications

(46 citation statements)

References 44 publications

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“…Metabolomics approaches may also be useful in optimizing media formulations. Miranda Yap and colleagues have utilized HPLC‐MS to characterize intracellular and extracellular metabolites derived from amino acids, media components, and various metabolic pathways, which can accumulate in the media during cell culture, inhibiting growth or triggering apoptosis . Additionally, Mulukutla and colleagues have used mass spectrometry and NMR to identify and quantitate metabolic byproducts, which were accumulating in a fed‐batch process and inhibiting growth, despite good control of lactate, ammonia and osmolality in the process .…”

Section: Recent Advances In Medium Developmentmentioning

confidence: 99%

Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Ritacco

Khetan

2018

Biotechnology Progress

192

138

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The culture of Chinese Hamster Ovary (CHO) cells for modern industrial applications, such as expression of recombinant proteins, requires media that support growth and production. Such media must support high viable cell densities while also stimulating the synthesis and extracellular transport of biologic products. Early media development efforts in this area yielded basic formulations to sustain growth, viability, and cellular function, albeit comprising animal sourced components, and complex constituents used in batch culture mode. Subsequent improvements included the development of serum‐free and chemically defined (CD) media, the identification of critical nutrients, growth factors, and potentially inhibitory or toxic cellular metabolites, and the use of fed‐batch and perfusion culture techniques to optimize nutrient delivery while minimizing accumulation of unwanted waste products. This review is comprised of sections covering milestones in the evolution of mammalian cell culture media, nutrient composition and formulation requirements, optimization strategies, consistency and scalability of powder and liquid media preparation for industrial applications, and key recent advances driving progress in CHO cell culture medium design and development. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1407–1426, 2018

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Section: Recent Advances In Medium Developmentmentioning

confidence: 99%

Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Ritacco

Khetan

2018

Biotechnology Progress

192

138

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“…Apoptosis is thought to be the main mechanism of cell death in industrial fed‐batch mammalian cell cultures due to stresses that arise by conditions that are used to maximize the manufacturing of therapeutic proteins (al‐Rubeai & Singh, ; Krampe & al‐Rubeai, ). Many groups have reported methods to reduce the rates of apoptosis as a means of extending culture viability, to increase protein production (Arden & Betenbaugh, ; Chong et al, ; Krampe & al‐Rubeai, ). A particularly successful example was the transfection of two inducible anti‐apoptotic genes into Chinese Hamster Ovary (CHO) cells that enabled increased fed‐batch viable cell concentrations as well as recombinant protein production (Figueroa et al, ).…”

Section: Introductionmentioning

confidence: 99%

Types of cell death and apoptotic stages in Chinese Hamster Ovary cells distinguished by Raman spectroscopy

Rangan

Kamal

Konorov

et al. 2017

Biotech & Bioengineering

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Cell death is the ultimate cause of productivity loss in bioreactors that are used to produce therapeutic proteins. We investigated the ability of Raman spectroscopy to detect the onset and types of cell death for Chinese Hamster Ovary (CHO) cells-the most widely used cell type for therapeutic protein production. Raman spectroscopy was used to compare apoptotic, necrotic, autophagic, and control CHO cells. Several specific nucleic acid-, protein-, and lipid-associated marker bands within the 650-850 cm spectral region were identified that distinguished among cells undergoing different modes of cell death; supporting evidence was provided by principal component analysis (PCA) of the full spectral data. In addition to comparing the different modes of cell death, normal cells were compared to cells sorted at several stages of apoptosis, in order to explore the potential for early detection of apoptosis. Different stages of apoptosis could be distinguished via Raman spectroscopy, with multiple changes observed in nucleic acid peaks at early stages whereas an increase in lipid signals was a feature of late apoptosis/secondary necrosis.

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“…Very early studies indicated a generalized reliance on glycolysis and glutaminolysis for energy production in early cell growth and information was available on other selected aspects of metabolism [9, 10]. However, recently, we have seen the optimization of analytical technologies that facilitate simultaneous characterization of multiple extracellular and intracellular metabolites for CHO cells in varied culture and environmental conditions [6, 11–15]. The mining and analysis of complex datasets from metabolite profiling (metabolomics approaches) has facilitated a greater understanding of the relationships between cells, medium, feeds and functional product generation [6, 15–17].…”

Section: Introductionmentioning

confidence: 99%

Metabolite profiling of CHO cells: Molecular reflections of bioprocessing effectiveness

Sellick

Croxford

Maqsood

et al. 2015

Biotechnology Journal

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Whilst development of medium and feeds has provided major advances in recombinant protein production in CHO cells, the fundamental understanding is limited. We have applied metabolite profiling with established robust (GC-MS) analytics to define the molecular loci by which two yield-enhancing feeds improve recombinant antibody yields from a model GS-CHO cell line. With data across core metabolic pathways, that report on metabolism within several cellular compartments, these data identify key metabolites and events associated with increased cell survival and specific productivity of cells. Of particular importance, increased process efficiency was linked to the functional activity of the mitochondria, with the amount and time course of use/production of intermediates of the citric acid cycle, for uses such as lipid biosynthesis, precursor generation and energy production, providing direct indicators of cellular status with respect to productivity. The data provide clear association between specific cellular metabolic indicators and cell process efficiency, extending from prior indications of the relevance of lactate metabolic balance to other redox sinks (glycerol, sorbitol and threitol). The information, and its interpretation, identifies targets for engineering cell culture efficiency, either from genetic or environmental perspectives, and greater understanding of the significance of specific medium components towards overall CHO cell bioprocessing.

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Metabolomics-based identification of apoptosis-inducing metabolites in recombinant fed-batch CHO culture media

Cited by 55 publications

References 44 publications

Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Cell culture media for recombinant protein expression in Chinese hamster ovary (CHO) cells: History, key components, and optimization strategies

Types of cell death and apoptotic stages in Chinese Hamster Ovary cells distinguished by Raman spectroscopy

Metabolite profiling of CHO cells: Molecular reflections of bioprocessing effectiveness

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