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Open AccessArticle

Chikungunya E2 Protein Produced in E. coli and HEK293-T Cells—Comparison of Their Performances in ELISA

1
Centro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, Brazil
2
Laboratório de Virologia Molecular e Aplicada, Departamento de Microbiologia, Instituto de Ciências Biológicas (ICB/UFMG), Belo Horizonte-MG 31270-901, Brazil
3
Colégio Técnico da Universidade Federal de Minas Gerais (COLTEC), Belo Horizonte-MG 31270-901, Brazil
4
The Jenner Institute, Nuffield Department of Medicine, The Henry Wellcome Building for Molecular Physiology, Roosevelt Drive, University of Oxford, Oxford OX3 7DQ, UK
*
Author to whom correspondence should be addressed.
Viruses 2020, 12(9), 939; https://doi.org/10.3390/v12090939
Received: 23 July 2020 / Revised: 18 August 2020 / Accepted: 19 August 2020 / Published: 26 August 2020
Chikungunya virus (CHIKV) is a mosquito-borne pathogen that causes a disease characterized by the acute onset of fever accompanied by arthralgia and intense joint pain. Clinical similarities and cocirculation of this and other arboviruses in many tropical countries highlight the necessity for efficient and accessible diagnostic tools. CHIKV envelope proteins are highly conserved among alphaviruses and, particularly, the envelope 2 glycoprotein (CHIKV-E2) appears to be immunodominant and has a considerable serodiagnosis potential. Here, we investigate how glycosylation of CHIKV-E2 affects antigen/antibody interaction and how this affects the performance of CHIKV-E2-based Indirect ELISA tests. We compare two CHIKV-E2 recombinant antigens produced in different expression systems: prokaryotic-versus eukaryotic-made recombinant proteins. CHIKV-E2 antigens are expressed either in E. coli BL21(DE3)—a prokaryotic system unable to produce post-translational modifications—or in HEK-293T mammalian cells—a eukaryotic system able to add post-translational modifications, including glycosylation sites. Both prokaryotic and eukaryotic recombinant CHIKV-E2 react strongly to anti-CHIKV IgG antibodies, showing accuracy levels that are higher than 90%. However, the glycan-added viral antigen presents better sensitivity and specificity (85 and 98%) than the non-glycosylated antigen (81 and 71%, respectively) in anti-CHIKV IgM ELISA assays. View Full-Text
Keywords: chikungunya virus; envelope protein 2; ELISA; heterologous expression; E. coli; HEK293-T cells chikungunya virus; envelope protein 2; ELISA; heterologous expression; E. coli; HEK293-T cells
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Bagno, F.F.; Godói, L.C.; Figueiredo, M.M.; Sérgio, S.A.R.; Moraes, T.F.S.; Salazar, N.C.; Kim, Y.C.; Reyes-Sandoval, A.; da Fonseca, F.G. Chikungunya E2 Protein Produced in E. coli and HEK293-T Cells—Comparison of Their Performances in ELISA. Viruses 2020, 12, 939.

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