Chikungunya E2 Protein Produced inE. coliand HEK293-T Cells-Comparison of Their Performances in ELISA

被引：14

作者：

Bagno, Flavia Fonseca ^{[1
,2
]}

Godoi, Lara Carvalho ^{[1
,3
]}

Figueiredo, Maria Marta ^{[1
]}

Rodrigues Sergio, Sarah Aparecida ^{[1
]}

Silva Moraes, Thais de Fatima ^{[1
,2
]}

Salazar, Natalia de Castro ^{[1
]}

Kim, Young Chan ^{[4
]}

Reyes-Sandoval, Arturo ^{[4
]}

da Fonseca, Flavio Guimaraes ^{[1
,2
]}

机构：

[1] Univ Fed Minas Gerais UFMG, Ctr Tecnol Vacinas CT Vacinas, Parque Tecnol UFMG BH Tec, BR-31320000 Belo Horizonte, MG, Brazil

[2] Inst Ciencias Biol ICB UFMG, Dept Microbiol, Lab Virol Mol & Aplicada, BR-31270901 Belo Horizonte, MG, Brazil

[3] Colegio Tecn Univ Fed Minas Gerais COLTEC, BR-31270901 Belo Horizonte, MG, Brazil

[4] Univ Oxford, Jenner Inst, Nuffield Dept Med, Henry Wellcome Bldg Mol Physiol, Roosevelt Dr, Oxford OX3 7DQ, England

来源：

VIRUSES-BASEL | 2020年 / 12卷 / 09期

基金：

“创新英国”项目;

关键词：

chikungunya virus; envelope protein 2; ELISA; heterologous expression; E; coli; HEK293-T cells; ENVELOPE PROTEINS; VIRUS; SERODIAGNOSIS; EXPRESSION; ENTRY; E1;

D O I：

10.3390/v12090939

中图分类号：

Q93 [微生物学];

学科分类号：

071005 [微生物学];

摘要：

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 inE. coliBL21(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.

引用

页数：12

共 41 条

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