Gas chromatography mass spectrometry (GCMS) - based global metabolite analysis of chinese hamster ovary (CHO) - K1 cells /
An efficient mammalian cell system for producing bioproducts should retain high cell viability and efficient use of energy sources rendering the need to understand the effects of various variables on the cell system.In this study, GCMS-based global metabolite analysis approach was used to study the...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
Kuala Lumpur :
Kulliyyah of Engineering, International Islamic University Malaysia,
2014
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| Subjects: | |
| Online Access: | Click here to view 1st 24 pages of the thesis. Members can view fulltext at the specified PCs in the library. |
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| Summary: | An efficient mammalian cell system for producing bioproducts should retain high cell viability and efficient use of energy sources rendering the need to understand the effects of various variables on the cell system.In this study, GCMS-based global metabolite analysis approach was used to study the relationships between growth phases (0 hour to 96 hour), types of media (RPMI 1640 and Ham's F12), level of medium composition (serum and glutamine) and types of microcarriers (Cytodex 1 and Cytodex 3) on culture growth behavior and productivity of CHO-K1 cells. CHO-KI cells producing IGF-1 protein were obtained from ATCC and grown in T-flask (37°C, 5% CO2) until 70-80% confluent in growth medium in T-flask and spinner flask. Samples were taken at 8-hourly intervals for routine cell counting (Tryphan Blue method), biochemical responses (glucose, glutamine and lactate concentration analyzed by YSI 2700), insulin like growth factor (IGF)-1 protein concentration (ELISA Kit, R&D Systems Inc.) and global metabolite analysis by GCMS. Conditioned media from each time point were spun down and collected before injected into GCMS. Data from GCMS was then transferred to SIMCA-P+12 for chemometric evaluation using principal component analysis (PCA). The results showed that while routine standard/ biochemical analysis gave only subtle differences between the media, GCMS-based global metabolite analysis was able to clearly separate the culture based on each type of condition aimed in this study. |
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| Physical Description: | xvii, 115 leaves : ill. ; 30cm. |
| Bibliography: | Includes bibliographical references (leaves 99-108) |