Immunogenicity and antigenic reactivity of a carrier-free synthetic peptide complex derived from a 70-kDaPlasmodium falciparum exoantigen

Abstract

A 70-kDaPlasmodium falciparum exoantigen was purified from supernatant fluids of continuous in vitroP. falciparum cultures using sequential cation-exchange and high performance liquid chromatographic procedures. The purified protein was then digested with chymotrypsin and amino-acid sequences were determined for the resulting fragments. Four peptides (termed C2, C3, C5, and C10) were subsequently selected for synthesis based on their predictability for antigenic sites. The peptides were effectively used as a synthetic immunogen (SPf70) when they had been copolymerized with glutaraldehyde in the absence of a carrier. When given with Freund's adjuvant, the synthetic peptide complex was found to be highly immunogenic in rabbits. Serologic reactivity to the peptide complex and peptides C2 and C5 was uniformly high, followed by the responses to peptides C3 and C10. Peptide antigenicity was also assessed with human anti-P. falciparum sera from malaria-endemic regions of Uganda and Venezuela. Enzymelinked immunosorbent assay (ELISA) data showed that anti-P. falciparum antibodies were specific for and reactive to the peptides. The specificity of the rabbit anti-SPf70 antibodies forP. falciparum antigen was shown by immunoprecipitation of metabolically labeled proteins and by immunoblotting. Herein we describe the peptide sequences of a 70-kDaP. falciparum exoantigen (Pf70) that, when synthesized and constructed as a copolymer (SPf70), are capable of inducing the formation of antibodies that are reactive with the native malarial protein. The high immunogenicity and antigenic reactivity of SPf70 indicate the potential use of this synthetic peptide polymer as an immunogen and a diagnostic reagent.