Effect of extracellular glutamine concentration on primary and secondary metabolism of a murine hybridoma: An in vivo13C nuclear magnetic resonance study

Mancuso, Anthony A.; Sharfstein, Susan T.; Fernandez, Erik J.; Clark, Douglas S.; Blanch, Harvey W.

doi:10.1002/(sici)1097-0290(19980120)57:2<172::aid-bit6>3.0.co;2-k

Cited by 25 publications

(6 citation statements)

References 31 publications

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“…A number of bioreactors inside spectrometers were designed at that time (see Section 3.7.). These permitted continuous monitoring of cell metabolism, notably to characterize the impact of drug-treatment and irradiation on mammalian cancer cell metabolism by 13 C-and 31 P-NMR [768,[777][778][779][780][781][782][783][784][785][786][787][788][789][790][791][792][793][794], or to study the effects of ethanol and various chemicals on hepatocytes [795,796], as well as metabolism in hybridoma [771,797]. However, in-cell NMR studies of metabolic activities appeared to be limited to cases involving the study of high concentration metabolites.…”

Section: The Early Days: Studying Carbon Fluxes Using 13 C-nmrmentioning

confidence: 99%

“…Soon afterwards in the mid-1980's, an advanced version of this principle was proposed to provide rapid flow of medium and high O 2 delivery for aerobic cells [1821,1822]. A similar approach involved circulating the fresh fluids through the lumen of hollow fibers, whose polymer porous walls permit nutrient diffusion to the interspace of the fibers, where suspension cells are maintained [771,797,1823]. Adherent cells can also attach on the hollow-fibers walls, either inside [1824] or outside [789,791,797,1823,[1825][1826][1827][1828][1829][1830].…”

Section: Cell Immobilization Strategiesmentioning

confidence: 99%

“…A similar approach involved circulating the fresh fluids through the lumen of hollow fibers, whose polymer porous walls permit nutrient diffusion to the interspace of the fibers, where suspension cells are maintained [771,797,1823]. Adherent cells can also attach on the hollow-fibers walls, either inside [1824] or outside [789,791,797,1823,[1825][1826][1827][1828][1829][1830]. Good results have also been obtained with a recent simplified version using a 1 MDa dialysis membrane, in which fresh medium flows continuously [1761].…”

Section: Cell Immobilization Strategiesmentioning

confidence: 99%

“…As discussed above (Section 2.3.2. ), in combination with 13 C-NMR, 31 P-NMR and bioreactors were used to study the effects of various drugs on mammalian cancer cell metabolism [768,[777][778][779][780][781][782][783][784][785][786][787][788][789][790][791][792][793][794], of ethanol and various chemicals on hepatocytes [795,796], and the metabolism of hybridoma [771,797], and yeast immobilized in gel matrices [866]. More recently, 31 P-NMR also provided interesting checks for cellular health during in-cell structural biology studies [867][868][869].…”

mentioning

confidence: 99%

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In-cell NMR: Why and how?

Theillet

Luchinat

2022

Progress in Nuclear Magnetic Resonance Spectroscopy

105

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Section: The Early Days: Studying Carbon Fluxes Using 13 C-nmrmentioning

confidence: 99%

Section: Cell Immobilization Strategiesmentioning

confidence: 99%

Section: Cell Immobilization Strategiesmentioning

confidence: 99%

mentioning

confidence: 99%

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In-cell NMR: Why and how?

Theillet

Luchinat

2022

Progress in Nuclear Magnetic Resonance Spectroscopy

105

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“…The nutrient feeding rate was kept equal to the nutrient consumption rates to maintain nutrients at low levels. Nutrient metabolism was then shifted to more efficient states [5,9–11]. Overall, the glucose and glutamine concentration was maintained at low levels, which caused a greater decrease in lactate and ammonia production rates when compared with that in BHK cells (80% versus 72%, and 65% versus 25% respectively).…”

Section: Discussionmentioning

confidence: 99%

Increasing the culture efficiency of hybridoma cells by the use of integrated metabolic control of glucose and glutamine at low levels

Feng

et al. 2005

Biotech and App Biochem

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The metabolism of HAb18 hybridoma cells was shifted to decrease metabolite accumulation and to improve culture efficiency by integrated metabolic control of glucose and glutamine at low levels. When glucose and glutamine levels were decreased to 0.5 and 0.3 mM respectively, lactate and ammonia production were reduced by 62.6 and 74% respectively, glucose-to-cell yield was increased from 0.23x10(9) to 0.66x10(9) cells.mmol-1, and glutamine-to-cell yield from 0.18x10(9) to 1.95x10(9) cells.mmol-1. Compared with high-level glucose and glutamine fed-batch cultures, low-level glucose and glutamine led to higher cell density (1.0x10(7) versus 0.3x10(7) cells.ml-1), longer culture span (14 as opposed to 8 days) and higher antibody yield (250 as against 150 mg.l-1). These results indicate that hybridoma culture efficiency would be increased by the integrated control of glucose and glutamine at 0.5 and 0.3 mM respectively. In contrast with previously reported glucose-and/or-glutamine-level-controlled fed-batch cultures, we demonstrated an efficient strategy of nutrient level selection and amino acid feeding. More importantly, our accurately and well-distributed Equable Feeding Control System opens a new avenue for reducing metabolites to low levels by controlling nutrients at low levels.

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Metabolic shifts by nutrient manipulation in continuous cultures of BHK cells

Cruz

Moreira

Carrondo

1999

Biotech & Bioengineering

114

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The present work aims at characterizing the regulatory mechanisms of metabolism and product formation of BHK cells producing a recombinant antibody/cytokine fusion protein. This work was carried out through the achievement of several steady‐states in chemostat cultures, corresponding to different glucose and glutamine levels in the feed culture medium. Results obtained indicate that both glucose and glutamine consumptions show a Michaelis–Menten dependence on residual glucose and glutamine concentrations, respectively. Similar dependence was also observed for lactate and ammonia productions. K GlcGlc and K GlnGln were estimated to be 0.4 and 0.15 mM, respectively, while q maxGlc and q maxGln were estimated to be 1.8 and 0.55 nmol 10−6cells min−1, respectively. At very low glucose concentrations, the glucose‐to‐lactate yield decreased markedly showing a metabolic shift towards lower lactate production; also, the glucose‐to‐cells yield was increased. At very low‐glutamine concentrations, the glutamine‐to‐ammonia and glutamine‐to‐cells yields increased, showing a more efficient glutamine metabolism. Overall, amino acid consumption was increased under low glucose or glutamine concentrations. Metabolic‐flux analysis confirmed the metabolic shifts by showing increases in the fluxes of the more energetically efficient pathways, at low‐nutrient concentrations. No effect of glucose or glutamine concentrations on the cell‐specific productivity was observed, even under metabolically shifted metabolism; therefore, it is possible to confine the cells to a more efficient metabolic state maintaining the productivity of the recombinant product of interest, and consequently, increasing final product titers by increasing cell concentration and culture length. This work is intended to be a model approach to characterize cell metabolism in an integrated way; it is highly valuable for the establishment of operating strategies in mammalian cell fermentations in which cell metabolism is to be confined to a desired state. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 66: 104–113, 1999.

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Effect of extracellular glutamine concentration on primary and secondary metabolism of a murine hybridoma: An in vivo13C nuclear magnetic resonance study

Cited by 25 publications

References 31 publications

In-cell NMR: Why and how?

In-cell NMR: Why and how?

Increasing the culture efficiency of hybridoma cells by the use of integrated metabolic control of glucose and glutamine at low levels

Metabolic shifts by nutrient manipulation in continuous cultures of BHK cells

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