Comparison between batch and fed-batch cultures of Chinese hamster ovary-DG44 cell using multi-well plate
The biopharmaceutical proteins, especially monoclonal antibodies have become of great significance in the drug manufacturing and development industry during the last two decades. This significance can be attributed to their effectiveness, selectivity and the wide range of diseases and health problem...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
2019
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| Subjects: | |
| Online Access: | http://eprints.utm.my/id/eprint/101992/1/AbdulKarimAbdulKaderMFS2019.pdf.pdf |
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| Summary: | The biopharmaceutical proteins, especially monoclonal antibodies have become of great significance in the drug manufacturing and development industry during the last two decades. This significance can be attributed to their effectiveness, selectivity and the wide range of diseases and health problems cured by them. Consequently, the demand for these proteins has increased greatly. To meet this increasing demand for these proteins, optimization and development of the production process is required. This optimization is done through intensive screening of different culture options which requires an effective screening tool. The traditional screening methods such as shake flask system are slow and cost-ineffective. Multi-well plate is an option to accelerate the process of development with a rational cost. The goal of this study is to evaluate the usage of the multi-well plate as a screening tool in the bioprocess development. To achieve this goal, 24-multi-well plate was used to compare between batch and fed-batch cultures of CHO-DG44 cells. Cell density, cell viability, cell morphology and confluency, glucose concentration, and total protein concentration were determined in both batch and fed-batch cultures. Cell counting was done using trypan blue excluding method with a haemocytometer, while the glucose concentration was determined using 3, 5-dinitrosalicylic acid (DNS) assay. On the other hand, total protein concentration was determined using Lowry assay. It has been observed that feeding nutrients during the fed-batch culture enhanced the cell density and viability compared to the batch culture. On day 7, the cell density and viability of the cells in batch culture were (3.667±1.04) ×104 cell/ml and (45.453±4.54) % respectively, while in fed-batch culture the cell density and viability were (5.167±0.76) ×104 cell/ml and (70.416±1.11) % respectively. Moreover, the protein production of the cells was prolonged during the fed-batch culture. This study revealed that the multi-well plate system can be used as a small-scale screening tool for optimization of CHO-cell culture. |
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