Construction and validation of Plasmodium antigens surface display library in mammalian cells.

• Main Author: Tan, Li Hua.
• Other Authors: School of Biological Sciences
• Format: Final Year Project
• Language: English
• Published: 2009
• Subjects: DRNTU::Science::Biological sciences::Microbiology::Immunology
• Online Access: http://hdl.handle.net/10356/16856
• Institution: Nanyang Technological University

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.