Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis

Mammalian cell lines, in particular CHO-K1 is vital for the multibillion dollar biotechnology industry. The majority of large scale bioprocessing of commercially aluable protein biopharmaceuticals is produced using this type of cell. An ideal mammalian cell system as host for biologics production sh...

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Main Authors:	Saberi, S.E.M., Hashim, Yumi Zuhanis Has-Yun, Mohamed, V.Packeer, Mel, Maizirwan, Ahmad Raus, Raha, Amid, Azura
Format:	Article
Language:	English
Published:	IIUM Press 2011
Subjects:	QR Microbiology
Online Access:	http://irep.iium.edu.my/17100/ http://irep.iium.edu.my/17100/ http://irep.iium.edu.my/17100/1/206.pdf

id	iium-17100
recordtype	eprints
spelling	iium-171002012-01-26T09:32:24Z http://irep.iium.edu.my/17100/ Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis Saberi, S.E.M. Hashim, Yumi Zuhanis Has-Yun Mohamed, V.Packeer Mel, Maizirwan Ahmad Raus, Raha Amid, Azura QR Microbiology Mammalian cell lines, in particular CHO-K1 is vital for the multibillion dollar biotechnology industry. The majority of large scale bioprocessing of commercially aluable protein biopharmaceuticals is produced using this type of cell. An ideal mammalian cell system as host for biologics production should retain efficient use of energy sources in order to boost productivity at minimum cost. Various analyses such as cell counting and monitoring of specific biochemical responses are used to provide data to enable bioprocess control in order to achieve the ideal system. Our study aimed to see whether global metabolite analysis using Gas Chromatography Mass Spectrometry (GCMS) would be a potential alternative approach in providing data for bioprocess control. In this study, we analyzed metabolites of CHO-K1 cells at different growth phases using GCMS. CHO-K1 cells producing insulin like growth factor-I (IGF1) were obtained from ATCC. Cells were grown in T-flask and incubated at 37°C/ 5% CO2 until 70-80% confluent in RPMI 1640 media. Samples (cells and spent/conditioned media) were taken at designated intervals for routine cell counting (Trypan Blue dye exclusion method); glucose, glutamine and lactate determination (YSI 2700); IGF-1 production (ELISA kit R&D Sstems, Inc); and global metabolite analysis (GCMS). Conditioned media from each time point were spun down before subjecting into GCMS. Data from GCMS was then transferred to SIMCA P+12.0 for chemometric evaluation using Principal Component Analysis (PCA). The first component, PC1 results was able to explain 36% of the variation of the data with clear separation between exponential phase and other phases (initial and death phase). This suggests that GCMS-based global metabolite analysis has the ability to capture cell growth behaviour and offered insights of factors that may influence the biological system. IIUM Press 2011 Article PeerReviewed application/pdf en http://irep.iium.edu.my/17100/1/206.pdf Saberi, S.E.M. and Hashim, Yumi Zuhanis Has-Yun and Mohamed, V.Packeer and Mel, Maizirwan and Ahmad Raus, Raha and Amid, Azura (2011) Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis. IIUM Engineering Journal, 12 (4). pp. 89-98. ISSN 1511-788X http://www.iium.edu.my/ejournal/index.php/iiumej/article/view/210/206
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	QR Microbiology
spellingShingle	QR Microbiology Saberi, S.E.M. Hashim, Yumi Zuhanis Has-Yun Mohamed, V.Packeer Mel, Maizirwan Ahmad Raus, Raha Amid, Azura Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis
description	Mammalian cell lines, in particular CHO-K1 is vital for the multibillion dollar biotechnology industry. The majority of large scale bioprocessing of commercially aluable protein biopharmaceuticals is produced using this type of cell. An ideal mammalian cell system as host for biologics production should retain efficient use of energy sources in order to boost productivity at minimum cost. Various analyses such as cell counting and monitoring of specific biochemical responses are used to provide data to enable bioprocess control in order to achieve the ideal system. Our study aimed to see whether global metabolite analysis using Gas Chromatography Mass Spectrometry (GCMS) would be a potential alternative approach in providing data for bioprocess control. In this study, we analyzed metabolites of CHO-K1 cells at different growth phases using GCMS. CHO-K1 cells producing insulin like growth factor-I (IGF1) were obtained from ATCC. Cells were grown in T-flask and incubated at 37°C/ 5% CO2 until 70-80% confluent in RPMI 1640 media. Samples (cells and spent/conditioned media) were taken at designated intervals for routine cell counting (Trypan Blue dye exclusion method); glucose, glutamine and lactate determination (YSI 2700); IGF-1 production (ELISA kit R&D Sstems, Inc); and global metabolite analysis (GCMS). Conditioned media from each time point were spun down before subjecting into GCMS. Data from GCMS was then transferred to SIMCA P+12.0 for chemometric evaluation using Principal Component Analysis (PCA). The first component, PC1 results was able to explain 36% of the variation of the data with clear separation between exponential phase and other phases (initial and death phase). This suggests that GCMS-based global metabolite analysis has the ability to capture cell growth behaviour and offered insights of factors that may influence the biological system.
format	Article
author	Saberi, S.E.M. Hashim, Yumi Zuhanis Has-Yun Mohamed, V.Packeer Mel, Maizirwan Ahmad Raus, Raha Amid, Azura
author_facet	Saberi, S.E.M. Hashim, Yumi Zuhanis Has-Yun Mohamed, V.Packeer Mel, Maizirwan Ahmad Raus, Raha Amid, Azura
author_sort	Saberi, S.E.M.
title	Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis
title_short	Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis
title_full	Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis
title_fullStr	Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis
title_full_unstemmed	Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis
title_sort	determination of igf-1-producing cho-k1 growth phases using gcms-based global metabolite analysis
publisher	IIUM Press
publishDate	2011
url	http://irep.iium.edu.my/17100/ http://irep.iium.edu.my/17100/ http://irep.iium.edu.my/17100/1/206.pdf
first_indexed	2023-09-18T20:25:54Z
last_indexed	2023-09-18T20:25:54Z
_version_	1777408414361583616

Determination of IGF-1-producing CHO-K1 growth phases using GCMS-based global metabolite analysis

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