Metabolomics-based identification of apoptosis-inducing metabolites in recombinant fed-batch CHO culture media

A liquid chromatography–mass spectrometry (LC–MS) based metabolomics platform was previously established to identify and profile extracellular metabolites in culture media of mammalian cells. This presented an opportunity to isolate novel apoptosis-inducing metabolites accumulating in the media of a...

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Bibliographic Details
Published in:Journal of biotechnology 2011-01, Vol.151 (2), p.218-224
Main Authors: Chong, William P.K., Yusufi, Faraaz N.K., Lee, Dong-Yup, Reddy, Satty G., Wong, Niki S.C., Heng, Chew Kiat, Yap, Miranda G.S., Ho, Ying Swan
Format: Article
Language:English
Subjects:
Adenosine - metabolism
Adenosine Triphosphate - metabolism
Animals
Apoptosis
Apoptosis Regulatory Proteins - chemistry
Biological and medical sciences
Bioreactors
Biotechnology
Caspases - metabolism
Cell Cycle
Chinese hamster ovary
CHO Cells
Chromatography, Liquid - methods
Cricetinae
Cricetulus
Fundamental and applied biological sciences. Psychology
Liquid chromatography–mass spectrometry
Mass Spectrometry - methods
Metabolic engineering
Metabolomics
Recombinant Proteins - chemistry
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Summary:A liquid chromatography–mass spectrometry (LC–MS) based metabolomics platform was previously established to identify and profile extracellular metabolites in culture media of mammalian cells. This presented an opportunity to isolate novel apoptosis-inducing metabolites accumulating in the media of antibody-producing Chinese hamster ovary (CHO mAb) fed-batch bioreactor cultures. Media from triplicate cultures were collected daily for the metabolomics analysis. Concurrently, cell pellets were obtained for determination of intracellular caspase activity. Metabolite profiles from the LC–MS data were subsequently examined for their degree of correlation with the caspase activity. A panel of extracellular metabolites, the majority of which were nucleotides/nucleosides and amino acid derivatives, exhibited good ( R 2 > 0.8) and reproducible correlation. Some of these metabolites, such as oxidized glutathione, AMP and GMP, were later shown to induce apoptosis when introduced to fresh CHO mAb cultures. Finally, metabolic engineering targets were proposed to potentially counter the harmful effects of these metabolites.
ISSN:0168-1656
1873-4863
DOI:10.1016/j.jbiotec.2010.12.010