Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines
The production of therapeutic monoclonal antibody is a time-consuming and costly process. With the increasing global demand for these recombinant proteins for therapeutic and research purposes, there is thus a need to improve this long and expensive production process. We have constructed several re...
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| Format: | Final Year Project (FYP) |
| Language: | English |
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2009
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| Online Access: | http://hdl.handle.net/10356/16320 |
| _version_ | 1826117453515587584 |
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| author | Lee, Eric Yong Jing |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Lee, Eric Yong Jing |
| author_sort | Lee, Eric Yong Jing |
| collection | NTU |
| description | The production of therapeutic monoclonal antibody is a time-consuming and costly process. With the increasing global demand for these recombinant proteins for therapeutic and research purposes, there is thus a need to improve this long and expensive production process. We have constructed several recombinant expression vectors to investigate the effects of selectable markers on the generation of high monoclonal antibody producing cell lines. The selectable markers investigated include neomycin-phosphotransferase (NPT) and its mutant variant which had reduced activity, dihydrofolate reductase (DHFR) and glutamine synthetase (GS). We stably selected for the cells and identified potential high-producers through surface fluorescent staining followed by fluorescent activated cell sorting. Due to time constraint, we could only assess the effect of wild-type NPT and its mutant variant. We observed that there were more potentially high-producing cells when the mutant NPT was used as compared to the wild-type NPT. This observation suggested that the mutation of the selectable marker sequence leading to its reduced activity had enhanced the stringency. As such, more high-titer cell clones were generated. Other selectable markers could be altered in a similar fashion to generate high-producing cell lines. |
| first_indexed | 2024-10-01T04:27:50Z |
| format | Final Year Project (FYP) |
| id | ntu-10356/16320 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:27:50Z |
| publishDate | 2009 |
| record_format | dspace |
| spelling | ntu-10356/163202023-02-28T18:06:24Z Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines Lee, Eric Yong Jing School of Biological Sciences Song, Zhi Wei DRNTU::Science::Biological sciences::Cytology The production of therapeutic monoclonal antibody is a time-consuming and costly process. With the increasing global demand for these recombinant proteins for therapeutic and research purposes, there is thus a need to improve this long and expensive production process. We have constructed several recombinant expression vectors to investigate the effects of selectable markers on the generation of high monoclonal antibody producing cell lines. The selectable markers investigated include neomycin-phosphotransferase (NPT) and its mutant variant which had reduced activity, dihydrofolate reductase (DHFR) and glutamine synthetase (GS). We stably selected for the cells and identified potential high-producers through surface fluorescent staining followed by fluorescent activated cell sorting. Due to time constraint, we could only assess the effect of wild-type NPT and its mutant variant. We observed that there were more potentially high-producing cells when the mutant NPT was used as compared to the wild-type NPT. This observation suggested that the mutation of the selectable marker sequence leading to its reduced activity had enhanced the stringency. As such, more high-titer cell clones were generated. Other selectable markers could be altered in a similar fashion to generate high-producing cell lines. Bachelor of Science in Biological Sciences 2009-05-25T04:14:03Z 2009-05-25T04:14:03Z 2009 2009 Final Year Project (FYP) http://hdl.handle.net/10356/16320 en Nanyang Technological University 33 p. application/pdf |
| spellingShingle | DRNTU::Science::Biological sciences::Cytology Lee, Eric Yong Jing Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines |
| title | Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines |
| title_full | Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines |
| title_fullStr | Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines |
| title_full_unstemmed | Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines |
| title_short | Effects of selectable markers on the generation of high monoclonal antibody-producing cell lines |
| title_sort | effects of selectable markers on the generation of high monoclonal antibody producing cell lines |
| topic | DRNTU::Science::Biological sciences::Cytology |
| url | http://hdl.handle.net/10356/16320 |
| work_keys_str_mv | AT leeericyongjing effectsofselectablemarkersonthegenerationofhighmonoclonalantibodyproducingcelllines |