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Fighting malaria is particularly challenging partly due to the complex life cycle of the parasite. However, technological breakthroughs such as the development of the nucleoside\u2010modified mRNA lipid nanoparticle (mRNA\u2010LNP) vaccine platform, along with the discovery of novel conserved <jats:italic>Plasmodium<\/jats:italic> antigens such as the E140 protein, present new opportunities in malaria prevention. Importantly, production of recombinant proteins for malaria vaccine evaluation by serological assays often represents an additional hurdle because many <jats:italic>Plasmodium<\/jats:italic> proteins are complex and often contain transmembrane domains that make production and purification particularly difficult. This research protocol provides a step\u2010by\u2010step guide for the production and purification of <jats:italic>P. berghei<\/jats:italic> and <jats:italic>P. vivax<\/jats:italic> E140 protein fragments that can be used to test humoral immune responses against this novel malaria vaccine target. We demonstrate that the purified proteins can be successfully used in enzyme\u2010linked immunosorbent assay (ELISA) to evaluate antigen\u2010specific binding antibody responses in sera obtained from E140 mRNA\u2010LNP\u2010vaccinated mice. Therefore, these proteins can contribute to the development and evaluation of E140\u2010based malaria vaccines.<\/jats:p>","DOI":"10.1002\/2211-5463.13939","type":"journal-article","created":{"date-parts":[[2025,1,16]],"date-time":"2025-01-16T06:04:38Z","timestamp":1737007478000},"page":"690-698","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Expression and purification of <scp>E140<\/scp> protein antigen fragments of <i>Plasmodium vivax<\/i> and <i>Plasmodium berghei<\/i> for serological assays"],"prefix":"10.1002","volume":"15","author":[{"given":"Rodolfo Ferreira","family":"Marques","sequence":"first","affiliation":[{"name":"Department of Microbiology Perelman School of Medicine, University of Pennsylvania  Philadelphia PA USA"},{"name":"Department of Parasitology Institute of Biomedical Sciences, University of Sao Paulo  S\u00e3o Paulo 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