Construction and validation of Plasmodium antigens surface display library in mammalian cells.
Malaria infection has been ongoing for the last 50,000 years, yet unlike other organisms, vaccine is still under-development. Experimental immunization with whole parasites live, physically or genetically attenuated provides successful protection in mice and humans. In order to identify the...
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| Format: | Final Year Project |
| Language: | English |
| Published: |
2009
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| Online Access: | http://hdl.handle.net/10356/16856 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | Malaria infection has been ongoing for the last 50,000 years, yet unlike other
organisms, vaccine is still under-development. Experimental immunization with
whole parasites live, physically or genetically attenuated provides successful
protection in mice and humans. In order to identify the antigens involved in efficient
immune response, we have developed tools and an assay to aid in the process of
identification. Malaria antigens from p.yoelii and p.falciparum were cloned and
expressed in mammalian cell surface display system. Using flow cytometry, nontransfected
and transfected cells were characterized by labeling with anti-myc and
the malaria fusion proteins were tested with anti-malaria antibodies. Similar
transfection rates were observed in the transfected HEK293 and HEK293T cells but
under drug pressure, only HEK293 cells could be used for positive transfectants
selection. Culturing of plasmid containing cells in 200μg/ml geneticin increased
percentage of cells containing plasmids. Transfected cells were specifically labeled
with anti-malaria antibodies, proving reliability of the assay for use in screening for
antigens recognized in immunized mice or humans. |
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