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Transient responses of hybridoma metabolism to changes in the oxygen supply rate in continuous culture

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Abstract

Oxygen is an important nutrient that may limit the productivity of commercial cell culture reactors. The transient responses of hybridoma growth and metabolism to step changes in the oxygen supply rate have been examined for dissolved oxygen concentrations (DO) ranging from 0.1% to 10% of air saturation in continuous culture. Metabolic quotients are reported for glucose, lactate, ammonia, oxygen, glutamine, alanine and other amino acids. A majority of the estimated ATP production was due to oxidative phosphorylation under all conditions tested. Decreases in the oxygen supply rate below the value required to maintain 0.5% DO caused the viable cell concentration to decrease. Glycolysis was enhanced at the lower oxygen concentrations, and after an initial decrease, the specific glutamine consumption rate was also higher. High residual glutamine concentrations occurred below 0.5% DO. Oxidation of other amino acids and production of serine were also inhibited. The cells subsequently adapted to low oxygen concentrations. The increase in cell concentration following the return to 10% DO was preceded by increased biosynthetic activity, as evidenced by transiently reduced yields of lactate from glucose, and alanine and ammonia from glutamine.

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Abbreviations

\(C_{{\text{O}}_2 } \) mM:

oxygen concentration

\(C_{_{{\text{O}}_2 } }^ * \) mM:

oxygen concentration in equilibrium with the headspace gas

D d−1 :

dilution rate = (volumetric feed rate)/(reactor volume)

f glycolysis mol/mol:

glycolysis correction factor = (glycolytic q ATP)/qlac

K L a h−1 :

volumetric mass transfer coefficient

n cells/cm3 :

total cell count

n v cells/cm3 :

viable cell count

p mM:

product concentration

p ATP mM cell−1 d−1 :

specific ATP production rate

\(Q_{{\text{O}}_2 } \) Mol/(m2 · h):

volumetric oxygen uptake rate

q g mM cell−1 d−1 :

specific glutamine consumption rate

\(q_{{\text{O}}_2 } \) mM cell−1 h−1 :

specific oxygen uptake rate

q p mM cell−1 d−1 :

specific production formation rate

q s mM cell−1 d−1 :

specific substrate consumption rate

P/O :

molecules of ATP generated per molecule of NADH oxidized

s mM:

substrate concentration

t d:

time

Y′ NADH, gln Mol/Mol:

mols of NADH (plus FADH2) oxidized per mol glutamine consumed

Y p,s Mol/Mol:

yield of product from substrate

Y′ p,s Mol/Mol:

apparent yield of product from sub strate = q p /p s

μ d−1 :

true specific growth rate

μ app d−1 :

apparent specific growth rate

ala:

alanine

NH3 :

ammonia

F :

feed stream

gluc:

glucose

glu:

glutamate

gln:

glutamine

lac:

lactate

NADH:

NADH plus FADH2

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  1. Department of Chemical Engineering, University of California, 94720, Berkeley, CA, USA

    W. M. Miller, C. R. Wilke & H. W. Blanch

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Miller, W.M., Wilke, C.R. & Blanch, H.W. Transient responses of hybridoma metabolism to changes in the oxygen supply rate in continuous culture. Bioprocess Engineering 3, 103–111 (1988). https://doi.org/10.1007/BF00373473

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