Computational design, expression, and characterization of a Plasmodium falciparum multi-epitope, multi-stage vaccine candidate (PfCTMAG).

Malaria, a tropical disease, claims the lives of thousands of people annually and the development of resistance to insecticides and antimalarial drugs poses a great challenge to current prevention and control strategies. Current malaria vaccines are limited in efficacy, duration of protection, and safety, due to the high antigenic diversity and complex life cycle of the Plasmodium parasite. This study sought to design and assess a more effective multi-stage, multi-epitope vaccine candidate for the control of malaria. A multi-epitope malaria vaccine candidate was designed in silico using multiple antigens from both the pre-erythrocytic and erythrocytic stages, expressed in bacteria, and its sero-reactivity to antibodies in plasma from malaria-positive (cases) and negative individuals (controls) was assessed using enzyme-linked immunosorbent assay (ELISA). Immunization experiments were equally conducted with BALB/c mice. In-silico analysis revealed that the designed antigen, PfCTMAG (Plasmodium falciparum Circumsporozoite, Thrombospondin-related adhesion protein, Merozoite surface protein 2, Apical asparagine (Asn)-rich protein and Glutamate-Rich Protein), effectively bound to Toll-like receptor 4 (TLR-4) and triggered a strong immune response. In sero-reactivity studies, malaria-positives (cases) had higher anti-PfCTMAG IgG ( p  = 0.024) and IgM ( p < 0.001) levels compared to malaria negatives (controls). The mice immunized with PfCTMAG did not show adverse reactions and had higher levels of IgG antibodies (p = 0.002) compared to controls, thereby validating the safety and immunogenicity of PfCTMAG as a promising vaccine candidate.