Marques, Rodolfo Ferreira and Ábrahám, Edit and Muramatsu, Hiromi and Bargieri, Daniel Youssef and Pardi, Norbert and Lipinszki, Zoltán (2025) Expression and purification of E140 protein antigen fragments of Plasmodium vivax and Plasmodium berghei for serological assays. FEBS OPEN BIO, 15 (5). pp. 690-698. ISSN 2211-5463
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Abstract
Malaria, a life-threatening disease caused by Plasmodium parasites, continues to pose a significant global health threat, with nearly 250 million infections and over 600 000 deaths reported annually by the WHO. 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-modified mRNA lipid nanoparticle (mRNA-LNP) vaccine platform, along with the discovery of novel conserved Plasmodium 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 Plasmodium proteins are complex and often contain transmembrane domains that make production and purification particularly difficult. This research protocol provides a step-by-step guide for the production and purification of P. berghei and P. vivax 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-linked immunosorbent assay (ELISA) to evaluate antigen-specific binding antibody responses in sera obtained from E140 mRNA-LNP-vaccinated mice. Therefore, these proteins can contribute to the development and evaluation of E140-based malaria vaccines.
| Item Type: | Article |
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| Additional Information: | Funding Agency and Grant Number: National Research, Development and Innovation Office (National Laboratory for Biotechnology) [2022-2.1.1-NL-2022-00008]; Hungarian Academy of Sciences [LP2017-7/2017]; National Research, Development, and Innovation Office, Hungary [2021-1.1.4-GYORSITOSAV-2022-00008]; Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2021/06769-0]; National Institute of Allergy and Infectious Diseases [R01AI146101, R01AI153064]; [2022/15895-2] Funding text: This work was supported by the National Research, Development and Innovation Office (National Laboratory for Biotechnology (2022-2.1.1-NL-2022-00008)) and the Hungarian Academy of Sciences (Lenduelet Program Grant (LP2017-7/2017)) to ZL and partially funded by the 2021-1.1.4-GYORSITOSAV-2022-00008 project of the National Research, Development, and Innovation Office, Hungary. RFM and DYB were supported by Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP 2021/06769-0 and 2022/15895-2, respectively). NP was supported by the National Institute of Allergy and Infectious Diseases (R01AI146101 and R01AI153064). We are grateful to Eva Boros (HUN-REN, BRC) for designing the graphical abstract. |
| Uncontrolled Keywords: | ELISA; protein purification; Malaria; serological assays; mRNA-lipid nanoparticle; <italic>Plasmodium</italic> spp.; recombinant E140 fragments; |
| Subjects: | Q Science / természettudomány > QR Microbiology / mikrobiológia |
| SWORD Depositor: | MTMT SWORD |
| Depositing User: | MTMT SWORD |
| Date Deposited: | 13 Mar 2026 08:09 |
| Last Modified: | 13 Mar 2026 08:09 |
| URI: | https://real.mtak.hu/id/eprint/235643 |
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