2023
DOI: 10.1002/biot.202200243
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Cottonseed hydrolysate supplementation alters metabolic and proteomics responses in Chinese hamster ovary cell cultures

Abstract: Hydrolysates are used as media supplements although their role is not well characterized. In this study, cottonseed hydrolysates, which contained peptides and galactose as supplemental substrates, were added to Chinese hamster ovary (CHO) batch cultures, enhancing cell growth, immunoglobulin (IgG) titers, and productivities. Extracellular metabolomics coupled with tandem mass tag (TMT) proteomics revealed metabolic and proteomic changes in cottonseed-supplemented cultures. Shifts in production and consumption … Show more

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Cited by 5 publications
(4 citation statements)
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“…There were numerous publications demonstrated the continuous supply of nutrients through the fed-batch possibly promoted glycolysis, aminoacyl-tRNA biosynthesis and TCA cycle, which might be considered to be responsible for increased mAb production. [36][37][38][39] The main pathways involved in the HS016 CHO-S cell culture process were also supported by these previous studies. Analysis of the correlated metabolic pathways of each phase was performed using the MetaboAnalyst web-server (http:// www.metaboanalyst.ca/MetaboAnalyst/) to characterize the important metabolic pathways.…”
Section: Analysis Of Metabolic Pathways For Three Cell Growth Phasessupporting
confidence: 74%
“…There were numerous publications demonstrated the continuous supply of nutrients through the fed-batch possibly promoted glycolysis, aminoacyl-tRNA biosynthesis and TCA cycle, which might be considered to be responsible for increased mAb production. [36][37][38][39] The main pathways involved in the HS016 CHO-S cell culture process were also supported by these previous studies. Analysis of the correlated metabolic pathways of each phase was performed using the MetaboAnalyst web-server (http:// www.metaboanalyst.ca/MetaboAnalyst/) to characterize the important metabolic pathways.…”
Section: Analysis Of Metabolic Pathways For Three Cell Growth Phasessupporting
confidence: 74%
“…Spent media collected were subjected to metabolite profiling using GC‐MS (Agilent 7890 GC with 5977 MSD). Extracellular concentrations of pyruvate, alanine, glycine, serine, aspartate, asparagine, and glutamate were measured using a GC‐MS‐based method used in previous studies (Dhara et al, 2023; Long & Antoniewicz, 2019; Y. Chen et al, 2019). Estimation of cell‐specific consumption rates ( q s ) were calculated using the following equation (Pan et al, 2017): Csfalse(tfalse)Csfalse(0false)=qsIVCD ${C}_{s}(t)-{C}_{s}(0)={q}_{s}{IVCD}$, where Csfalse(tfalse) ${C}_{s}(t)$ is the concentration of substrate S at time t, Csfalse(0false) ${C}_{s}(0)$ is the concentration of substrate S at the start of the culture.…”
Section: Methodsmentioning
confidence: 99%
“…Extracellular concentrations of pyruvate, alanine, glycine, serine, aspartate, asparagine, and glutamate were measured using a GC-MSbased method used in previous studies (Dhara et al, 2023;Long & Antoniewicz, 2019;. Estimation of cell-specific consumption rates (q s ) were calculated using the following equation (Pan et al, 2017):…”
Section: Extracellular and Intracellular Metabolite Profilingmentioning
confidence: 99%
“…This continued until both the nutrients in the culture were completely exhausted leading to the stopping of cell growth and subsequently cell death.Typically, commercial media are designed to sustain growth for a wide range of CHO cell lines and thus contain relatively standard amounts of different nutrients utilized as part of normal cellular metabolism. Knowledge about the amino acid concentrations over the duration of the batch culture will provide us with useful information about amino acid consumption patterns of the CHO cells in culture(Dhara et al, 2023). Therefore, shown in Figure2is a heat map profile of relative levels of amino acids in CHO cells (normalized to 100%) over time for a batch cell culture process using medium A supplemented with 6 mM L-Glutamine.…”
mentioning
confidence: 99%