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Dengue and Zika virus multi-epitope antigen expression in insect cells

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Abstract

Dengue virus and Zika virus are arthropod-borne flaviviruses that cause millions of infections worldwide. The co-circulation of both viruses makes serological diagnosis difficult as they share high amino acid similarities in viral proteins. Antigens are one of the key reagents in the differential diagnosis of these viruses through the detection of IgG antibodies in serological assays during the convalescent-phase of infections. Here, we report the expression of Dengue virus (DENV) and Zika virus (ZIKV) antigens containing non-conserved and immunodominant amino acid sequences using the baculovirus expression vector system in insect cells. We designed DENV and ZIKV antigens based on the domain III of the E protein (EDIII) after analyzing previously reported epitopes and by multiple alignment of the most important flaviviruses. The ZIKV and DENV multi-epitope genes were designed as tandem repeats or impaired repeats separated by tetra- or hexa-glycine linkers. The biochemical analyses revealed adequate expression of the antigens. Then, the obtained multi-epitope antigens were semi-purified in a sucrose gradient and tested using patients’ sera collected during the convalescent-phase that were previously diagnosed positive for anti-DENV and -ZIKV IgG antibodies. The optimal serum dilution was 1:200, and the mean absorbance values in the preliminary tests show that multi-epitope antigens have been recognized by human sera. The production of both antigens using the multi-epitope strategy in the eukaryotic system and based on the EDIII regions provide a proof of concept for the use of antigens in the differentiation between DENV and ZIKV.

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Acknowledgements

This work was supported by grants from Fundação de Apoio a Pesquisa do Distrito Federal (FAP/DF Grant Numbers 193.001532/2016 and 0193.000416/2016) and was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001". Our thanks to the researchers of the “Zika Plan” group (734584-European Union’s Horizon 2020) and also to Apoio a Pesquisa do Estado de Goiás (FAPEG) (Project Number 201710267001241) for financial support.

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Authors and Affiliations

  1. Campus Colemar Natal E Silva, Instituto de Patologia Tropical E Saúde Pública, Universidade Federal de Goiás, Goiânia, GO, 74605-450, Brazil

    Leonardo Lopes-Luz & Lucimeire Antonelli da Silveira

  2. Faculdade de Farmácia, Campus Colemar Natal E Silva, Universidade Federal de Goiás, Goiânia, GO, 74605-170, Brazil

    Isabela Cinquini Junqueira

  3. Departamento de Biologia Celular, Campus Darcy Ribeiro, Universidade de Brasília, Brasília, DF, 70910-900, Brazil

    Bruna Ribeiro de Melo Pereira, Leonardo Assis da Silva, Bergmann Morais Ribeiro & Tatsuya Nagata

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  1. Leonardo Lopes-Luz
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Correspondence to Tatsuya Nagata.

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Lopes-Luz, L., Junqueira, I.C., da Silveira, L.A. et al. Dengue and Zika virus multi-epitope antigen expression in insect cells. Mol Biol Rep 47, 7333–7340 (2020). https://doi.org/10.1007/s11033-020-05772-1

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