Production And Characterization Of Monoclonal Antibodies Against Recombinate Malaria Vaccine PfCP-2.9

Malaria remains one of the most devasting infectious diseases in the world. Due to emergence of the drug resistance of the parasite and the insecticide resistance of Anopheles mosquito, it becomes more difficult to control malaria. So it is very important to develop effective malaria vaccine to control the disease.Since only asexual stage of the plasmodium falciparum 's life cycle causes disease, development of malaria vaccine against this stage should reduce the mortality and morbidity of the disease. Two antigens, the 19kDa carboxyl-terminai damain of Merozoite Surface Protein 1 (MSPl-19) and the domain III -of Apical Membrane Antigen 1 [AMA-1(III)] have been considered a leading vaccine candidates because of experimental evidences showing that they can induce inhibitory antibodies. A chimeric protein named PfCP-2.9 was constructed in this laboratory, which is composed of MSPl-19 and AMA-1(III) of plasmodium falciparum via a hinge sequence. As a malaria vaccine, PfCP-2.9 has been admitted to have clinical trial and phase I has been finished. The chimeric protein PfCP-2.9 is highly immunogenic, and specific antibody titer is 10 and 18 times than MSPl-19 and AMA-1(III). The animal immune serum with PfCP-2.9 strongly inhibited parasite growth, while MSPl-19 and AMA-1(III) did not when concentration being 15%. The serum of malaria sufferer also have higher immunoreactivity with PfCP-2.9 than with MSPl-19 and AMA-1(III). All above indicated that MSPl-19 and AMA-1(III)'s immunogenicity, immunoreactivity and inhibitory efficiency be enhanced after being fused.To approach the mechanism of the enhancement of PfCP-2.9's immunogenicity and inhibitory efficiency, anti-PfCP-2.9 monoclonal antibodies were generated and investigated. Among the epitopes which recognized by these mAbs, the inhibitory ones and the new ones were analyzed to find the the mechanism of the enhancement of PfCP-2.9's immunogenicity and inhibitory efficiency, and the works will also provide some idea about the vaccine design.In our work, BALB/c mice were immunized with PfCP-2.9, and then it's spleen cells were used to fusion with SP2/0 cells. There are 20 mAbs be gained via ELISA...