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

Result code in 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>

Result on the web

<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>

Result language

angličtina

Original language name

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

Original language description

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.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

20401 - Chemical engineering (plants, products)

Project

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Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2017

Confidentiality

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

Name of the periodical

Biotechnology Progress

ISSN

1520-6033

e-ISSN

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Volume of the periodical

33

Issue of the periodical within the volume

6

Country of publishing house

US - UNITED STATES

Number of pages

10

Pages from-to

"1630?1639"

UT code for WoS article

000418802100022

EID of the result in the Scopus database

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