Cookies on this website

We use cookies to ensure that we give you the best experience on our website. If you click 'Accept all cookies' we'll assume that you are happy to receive all cookies and you won't see this message again. If you click 'Reject all non-essential cookies' only necessary cookies providing core functionality such as security, network management, and accessibility will be enabled. Click 'Find out more' for information on how to change your cookie settings.

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.

Original publication

DOI

10.3390/v12090939

Type

Journal article

Journal

Viruses

Publication Date

26/08/2020

Volume

12

Addresses

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.

Keywords

Humans, Escherichia coli, Chikungunya virus, Polysaccharides, Immunoglobulin G, Immunoglobulin M, Recombinant Proteins, Viral Envelope Proteins, Antibodies, Viral, Antigens, Viral, Enzyme-Linked Immunosorbent Assay, Serologic Tests, Sensitivity and Specificity, Glycosylation, HEK293 Cells, Chikungunya Fever