Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISA
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 effici...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-08-01
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| Series: | Viruses |
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| Online Access: | https://www.mdpi.com/1999-4915/12/9/939 |
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| author | Flávia Fonseca Bagno Lara Carvalho Godói Maria Marta Figueiredo Sarah Aparecida Rodrigues Sérgio Thaís de Fátima Silva Moraes Natália de Castro Salazar Young Chan Kim Arturo Reyes-Sandoval Flávio Guimarães da Fonseca |
| author_facet | Flávia Fonseca Bagno Lara Carvalho Godói Maria Marta Figueiredo Sarah Aparecida Rodrigues Sérgio Thaís de Fátima Silva Moraes Natália de Castro Salazar Young Chan Kim Arturo Reyes-Sandoval Flávio Guimarães da Fonseca |
| author_sort | Flávia Fonseca Bagno |
| collection | DOAJ |
| description | 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 <i>E. coli</i> 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. |
| first_indexed | 2024-03-10T16:49:22Z |
| format | Article |
| id | doaj.art-ffdbbf0b71324bf5837bb5b0c5fb4215 |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T16:49:22Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-ffdbbf0b71324bf5837bb5b0c5fb42152023-11-20T11:22:03ZengMDPI AGViruses1999-49152020-08-0112993910.3390/v12090939Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISAFlávia Fonseca Bagno0Lara Carvalho Godói1Maria Marta Figueiredo2Sarah Aparecida Rodrigues Sérgio3Thaís de Fátima Silva Moraes4Natália de Castro Salazar5Young Chan Kim6Arturo Reyes-Sandoval7Flávio Guimarães da Fonseca8Centro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, BrazilCentro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, BrazilCentro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, BrazilCentro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, BrazilCentro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, BrazilCentro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, BrazilThe Jenner Institute, Nuffield Department of Medicine, The Henry Wellcome Building for Molecular Physiology, Roosevelt Drive, University of Oxford, Oxford OX3 7DQ, UKThe Jenner Institute, Nuffield Department of Medicine, The Henry Wellcome Building for Molecular Physiology, Roosevelt Drive, University of Oxford, Oxford OX3 7DQ, UKCentro de Tecnologia em Vacinas (CT-Vacinas), Parque Tecnológico da UFMG (BH-Tec), Universidade Federal de Minas Gerais (UFMG), Belo Horizonte-MG 31320-000, BrazilChikungunya 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 <i>E. coli</i> 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.https://www.mdpi.com/1999-4915/12/9/939chikungunya virusenvelope protein 2ELISAheterologous expression<i>E. coli</i>HEK293-T cells |
| spellingShingle | Flávia Fonseca Bagno Lara Carvalho Godói Maria Marta Figueiredo Sarah Aparecida Rodrigues Sérgio Thaís de Fátima Silva Moraes Natália de Castro Salazar Young Chan Kim Arturo Reyes-Sandoval Flávio Guimarães da Fonseca Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISA Viruses chikungunya virus envelope protein 2 ELISA heterologous expression <i>E. coli</i> HEK293-T cells |
| title | Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISA |
| title_full | Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISA |
| title_fullStr | Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISA |
| title_full_unstemmed | Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISA |
| title_short | Chikungunya E2 Protein Produced in <i>E. coli</i> and HEK293-T Cells—Comparison of Their Performances in ELISA |
| title_sort | chikungunya e2 protein produced in i e coli i and hek293 t cells comparison of their performances in elisa |
| topic | chikungunya virus envelope protein 2 ELISA heterologous expression <i>E. coli</i> HEK293-T cells |
| url | https://www.mdpi.com/1999-4915/12/9/939 |
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