Isotope labeling to determine the dynamics of metabolic response in CHO cell perfusion cultures using MALDI-TOF-MS

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60461373%3A22340%2F17%3A43914795" target="_blank" >RIV/60461373:22340/17:43914795 - isvavai.cz</a>

Výsledek na webu

<a href="http://onlinelibrary.wiley.com/doi/10.1002/btpr.2539/abstract" target="_blank" >http://onlinelibrary.wiley.com/doi/10.1002/btpr.2539/abstract</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/btpr.2539" target="_blank" >10.1002/btpr.2539</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Isotope labeling to determine the dynamics of metabolic response in CHO cell perfusion cultures using MALDI-TOF-MS

Popis výsledku v původním jazyce

The steady-state operation of Chinese hamster ovary (CHO) cells in perfusion bioreactors requires the equilibration of reactor dynamics and cell metabolism. Accordingly, in this work we investigate the transient cellular response to changes in its environment and their interactions with the bioreactor hydrodynamics. This is done in a benchtop perfusion bioreactor using MALDI-TOF MS through isotope labeling of complex intracellular nucleotides (ATP, UTP) and nucleotide sugars (UDP-Hex, UDP-HexNAc). By switching to a 13C6 glucose containing feed media during constant operation at 20 × 106 cells and a perfusion rate of 1 reactor volume per day, isotopic steady state was studied. A step change to the 13C6 glucose medium in spin tubes allowed the determination of characteristic times for the intracellular turnover of unlabeled metabolites pools, math formula (?0.56 days), which were confirmed in the bioreactor. On the other hand, it is shown that the reactor residence time math formula (1 day) and characteristic time for glucose uptake math formula (0.33 days), representative of the bioreactor dynamics, delayed the consumption of 13C6 glucose in the bioreactor and thus the intracellular 13C enrichment. The proposed experimental approach allowed the decoupling of bioreactor hydrodynamics and intrinsic dynamics of cell metabolism in response to a change in the cell culture environment.

Název v anglickém jazyce

Isotope labeling to determine the dynamics of metabolic response in CHO cell perfusion cultures using MALDI-TOF-MS

Popis výsledku anglicky

The steady-state operation of Chinese hamster ovary (CHO) cells in perfusion bioreactors requires the equilibration of reactor dynamics and cell metabolism. Accordingly, in this work we investigate the transient cellular response to changes in its environment and their interactions with the bioreactor hydrodynamics. This is done in a benchtop perfusion bioreactor using MALDI-TOF MS through isotope labeling of complex intracellular nucleotides (ATP, UTP) and nucleotide sugars (UDP-Hex, UDP-HexNAc). By switching to a 13C6 glucose containing feed media during constant operation at 20 × 106 cells and a perfusion rate of 1 reactor volume per day, isotopic steady state was studied. A step change to the 13C6 glucose medium in spin tubes allowed the determination of characteristic times for the intracellular turnover of unlabeled metabolites pools, math formula (?0.56 days), which were confirmed in the bioreactor. On the other hand, it is shown that the reactor residence time math formula (1 day) and characteristic time for glucose uptake math formula (0.33 days), representative of the bioreactor dynamics, delayed the consumption of 13C6 glucose in the bioreactor and thus the intracellular 13C enrichment. The proposed experimental approach allowed the decoupling of bioreactor hydrodynamics and intrinsic dynamics of cell metabolism in response to a change in the cell culture environment.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

20401 - Chemical engineering (plants, products)

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Biotechnology Progress

ISSN

1520-6033

e-ISSN

—

Svazek periodika

33

Číslo periodika v rámci svazku

6

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

10

Strana od-do

"1630?1639"

Kód UT WoS článku

000418802100022

EID výsledku v databázi Scopus

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