Preparing domain Ⅱ F2 fragment of Plasmodium falciparum 175000 erythrocyte binding antigen and its combined immunization / 第二军医大学学报

ABSTRACT

Objective:To synthesize the eba 175Ⅱf2 gene of Plasmodium falciparum and express it in Pichia pastoris in the secreting form. The recombinant protein and PfCP 2.9 protein were used for combined immunization to see if there is antigen competition. Methods: Asymmetric PCR based metho d was utilized to synthesize the 959bp eba 175Ⅱf2 gene. Plasmid containing the synthetic gene was introduced into Pichi a pa storis by electroporation for inducible expression. The recombinant EBA 175Ⅱ F2 p rotein was purified by ion exchange and gel filtration ch romatography. Results: The eba 175Ⅱf2 gene was successfully ex p ressed in Pichia pastoris in the secreting form. The antibody titers in mice immunized with the combined EBA 175ⅡF2 and PfCP 2.9 proteins w ere much higher tha n those with individual protein by ELISA. Conclusion: No antigen competition is found when using the combined EBA 175ⅡF2 and PfCP 2.9 immunization in mice, indicating the potential of combining the 2 antigens for vaccination.