Expression and purification of allelic variants of Plasmodium vivax Apical Membrane Antigen 1 (AMA-1) aiming to immunological studies

Abstract

Malaria is one of the priorities of world research in the field of vaccine development, with Plasmodium vivax being the most geographically distributed species in the world and the most prevalent species in the Americas. Unlike Plasmodium falciparum, there is currently no vaccine against P. vivax in advanced clinical trials. Although efforts to design vaccines against P. vivax have been important, one limitation is the presence of antigenic polymorphisms, a crucial factor for most candidate vaccines such as PvAMA1. To eliminate this gap we intend to investigate the presence of polymorphisms in 4 allelic variants of PvAMA1. The objective is to express and purify the recombinant proteins representing the following allelic forms of P. vivax AMA1: PvAMA-PNG_62_MU, PvAMA-TC103, PvAMA-Chesson-I and PvAMA-PNG_5. For this, the allelic variants of PvAMA1 will be expressed in Pichia pastoris yeasts and purified by affinity chromatography and ion exchange. Its detection will be confirmed by immunoblotting and the purity by high performance chromatography (HPLC). These data in the future will be valuable for the study of the impact of PvAMA1 polymorphism on recognition by specific antibodies, aiming to contribute to the development of a global multiple effective allele vaccine against P. vivax malaria. (AU)