Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine

Background: Gas Chromatography Mass Spectrometry (GCMS)-based metabolomics has been shown to substantially contribute to understanding of the physiological state of mammalian producer cells leading to improved large-scale production of biopharmaceuticals. In this study, GCMSbased metabolomics (gl...

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Main Authors:	Hashim, Yumi Zuhanis Has-Yun, Mohmad Saberi, Salfarin Ezrina, Mel, Maizirwan, Packeer Mohamed, Vasila
Format:	Article
Language:	English English
Published:	Bentham Science 2016
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/47164/ http://irep.iium.edu.my/47164/ http://irep.iium.edu.my/47164/ http://irep.iium.edu.my/47164/1/Paper-Hashim_et_al_%282016%29_CMB_.pdf http://irep.iium.edu.my/47164/4/56444_Profiling%20of%20Extracellular_WOS.pdf

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spelling	iium-471642017-04-12T06:45:31Z http://irep.iium.edu.my/47164/ Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine Hashim, Yumi Zuhanis Has-Yun Mohmad Saberi, Salfarin Ezrina Mel, Maizirwan Packeer Mohamed, Vasila Q Science (General) Background: Gas Chromatography Mass Spectrometry (GCMS)-based metabolomics has been shown to substantially contribute to understanding of the physiological state of mammalian producer cells leading to improved large-scale production of biopharmaceuticals. In this study, GCMSbased metabolomics (global metabolite analysis) approach was used to understand the effects of different levels of serum and glutamine on metabolite profiles associated with cell growth behaviour and insulin-like growth factor 1 (IGF1) protein production. Methods: CHO-KI cells producing IGF1 were obtained from American Type Culture Collection (ATCC) and grown in Tflask (37°C, 5 % CO2) until 70-80 % confluent in optimized medium, RPMI 1640 with different levels of of serum (%) and glutamine (mM). The different compositions of serum and glutamine were based on three levels full factorial design generated by MODDE, SIMCA P+Version 12 (Umetrics). Samples were then taken at 8-hourly intervals for routine cell counting, biochemical responses, IGF1 protein concentration and global metabolite analysis (GCMS). Conditioned media from each time point were spun down before injection into GCMS. Data from GCMS were then transferred to SIMCAP+ Version 12 (Umetrics) for chemometric evaluation using Partial Least Square Discriminant Analysis (PLS-DA). Results: Experiment Run 3 which produced the highest cell density was discriminated from other runs based on the extracellular metabolites. This indicates that Run 3 although supplied with very little glutamine, was able to undergo active metabolism including glycolysis and as such has efficiently used up the nutrient sources to produce the highest cell number. However, no clear relationship can be delineated for the IGF1 production. Conclusion: Global metabolite analysis approach has been proven to be able to give more insights into the metabolism of cells as compared to routine biochemical studies where data was less informative. Bentham Science 2016-04 Article PeerReviewed application/pdf en http://irep.iium.edu.my/47164/1/Paper-Hashim_et_al_%282016%29_CMB_.pdf application/pdf en http://irep.iium.edu.my/47164/4/56444_Profiling%20of%20Extracellular_WOS.pdf Hashim, Yumi Zuhanis Has-Yun and Mohmad Saberi, Salfarin Ezrina and Mel, Maizirwan and Packeer Mohamed, Vasila (2016) Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine. Current Metabolomics, 4 (1). pp. 56-62. ISSN 2213-235X E-ISSN 2213-2368 http://benthamscience.com/journals/current-metabolomics/article/135048/ 10.2174/2213235X03666150917183658
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English
topic	Q Science (General)
spellingShingle	Q Science (General) Hashim, Yumi Zuhanis Has-Yun Mohmad Saberi, Salfarin Ezrina Mel, Maizirwan Packeer Mohamed, Vasila Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine
description	Background: Gas Chromatography Mass Spectrometry (GCMS)-based metabolomics has been shown to substantially contribute to understanding of the physiological state of mammalian producer cells leading to improved large-scale production of biopharmaceuticals. In this study, GCMSbased metabolomics (global metabolite analysis) approach was used to understand the effects of different levels of serum and glutamine on metabolite profiles associated with cell growth behaviour and insulin-like growth factor 1 (IGF1) protein production. Methods: CHO-KI cells producing IGF1 were obtained from American Type Culture Collection (ATCC) and grown in Tflask (37°C, 5 % CO2) until 70-80 % confluent in optimized medium, RPMI 1640 with different levels of of serum (%) and glutamine (mM). The different compositions of serum and glutamine were based on three levels full factorial design generated by MODDE, SIMCA P+Version 12 (Umetrics). Samples were then taken at 8-hourly intervals for routine cell counting, biochemical responses, IGF1 protein concentration and global metabolite analysis (GCMS). Conditioned media from each time point were spun down before injection into GCMS. Data from GCMS were then transferred to SIMCAP+ Version 12 (Umetrics) for chemometric evaluation using Partial Least Square Discriminant Analysis (PLS-DA). Results: Experiment Run 3 which produced the highest cell density was discriminated from other runs based on the extracellular metabolites. This indicates that Run 3 although supplied with very little glutamine, was able to undergo active metabolism including glycolysis and as such has efficiently used up the nutrient sources to produce the highest cell number. However, no clear relationship can be delineated for the IGF1 production. Conclusion: Global metabolite analysis approach has been proven to be able to give more insights into the metabolism of cells as compared to routine biochemical studies where data was less informative.
format	Article
author	Hashim, Yumi Zuhanis Has-Yun Mohmad Saberi, Salfarin Ezrina Mel, Maizirwan Packeer Mohamed, Vasila
author_facet	Hashim, Yumi Zuhanis Has-Yun Mohmad Saberi, Salfarin Ezrina Mel, Maizirwan Packeer Mohamed, Vasila
author_sort	Hashim, Yumi Zuhanis Has-Yun
title	Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine
title_short	Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine
title_full	Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine
title_fullStr	Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine
title_full_unstemmed	Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine
title_sort	profiling of extracellular metabolites in cho-k1 cells cultured in medium with different levels of serum and glutamine
publisher	Bentham Science
publishDate	2016
url	http://irep.iium.edu.my/47164/ http://irep.iium.edu.my/47164/ http://irep.iium.edu.my/47164/ http://irep.iium.edu.my/47164/1/Paper-Hashim_et_al_%282016%29_CMB_.pdf http://irep.iium.edu.my/47164/4/56444_Profiling%20of%20Extracellular_WOS.pdf
first_indexed	2023-09-18T21:07:08Z
last_indexed	2023-09-18T21:07:08Z
_version_	1777411008681213952

Profiling of extracellular metabolites in CHO-K1 cells cultured in medium with different levels of serum and glutamine

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