Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs

Autores
Serena, Maria Soledad; Geisler, Christoph; Metz, German Ernesto; Corva, Santiago Gerardo; Mórtola, Eduardo Carlos; Larsen, Alejandra; Jarvis, Donald L.; Echeverria, Maria Gabriela
Año de publicación
2013
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
Fil: Serena, Maria Soledad. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Geisler, Christoph. University of Wyoming. Department of Molecular Biology; Estados Unidos
Fil: Metz, German Ernesto. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Corva, Santiago Gerardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Mórtola, Eduardo Carlos. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Jarvis, Donald L.. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Echeverria, Maria Gabriela. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
Suid Herpesvirus 1
Baculovirus
Insect Cell
Glycoprotein
Elisa
Gc Content
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/11596
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/11596
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigsSerena, Maria SoledadGeisler, ChristophMetz, German ErnestoCorva, Santiago GerardoMórtola, Eduardo CarlosLarsen, AlejandraJarvis, Donald L.Echeverria, Maria GabrielaSuid Herpesvirus 1BaculovirusInsect CellGlycoproteinElisaGc Contenthttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.Fil: Serena, Maria Soledad. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Geisler, Christoph. University of Wyoming. Department of Molecular Biology; Estados UnidosFil: Metz, German Ernesto. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Corva, Santiago Gerardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Mórtola, Eduardo Carlos. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Jarvis, Donald L.. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; ArgentinaFil: Echeverria, Maria Gabriela. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaElsevier2013-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/11596Serena, Maria Soledad; Geisler, Christoph; Metz, German Ernesto; Corva, Santiago Gerardo; Mórtola, Eduardo Carlos; et al.; Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs; Elsevier; Protein Expression And Purification; 90; 1; 7-2013; 1-81046-5928enginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2013.04.008info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1046592813000752info:eu-repo/semantics/altIdentifier/url/https://pubmed.ncbi.nlm.nih.gov/23631926/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:40:25Zoai:ri.conicet.gov.ar:11336/11596instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:40:26.131CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
title Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
spellingShingle Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
Serena, Maria Soledad
Suid Herpesvirus 1
Baculovirus
Insect Cell
Glycoprotein
Elisa
Gc Content
title_short Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
title_full Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
title_fullStr Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
title_full_unstemmed Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
title_sort Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs
dc.creator.none.fl_str_mv Serena, Maria Soledad
Geisler, Christoph
Metz, German Ernesto
Corva, Santiago Gerardo
Mórtola, Eduardo Carlos
Larsen, Alejandra
Jarvis, Donald L.
Echeverria, Maria Gabriela
author Serena, Maria Soledad
author_facet Serena, Maria Soledad
Geisler, Christoph
Metz, German Ernesto
Corva, Santiago Gerardo
Mórtola, Eduardo Carlos
Larsen, Alejandra
Jarvis, Donald L.
Echeverria, Maria Gabriela
author_role author
author2 Geisler, Christoph
Metz, German Ernesto
Corva, Santiago Gerardo
Mórtola, Eduardo Carlos
Larsen, Alejandra
Jarvis, Donald L.
Echeverria, Maria Gabriela
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Suid Herpesvirus 1
Baculovirus
Insect Cell
Glycoprotein
Elisa
Gc Content
topic Suid Herpesvirus 1
Baculovirus
Insect Cell
Glycoprotein
Elisa
Gc Content
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
Fil: Serena, Maria Soledad. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Geisler, Christoph. University of Wyoming. Department of Molecular Biology; Estados Unidos
Fil: Metz, German Ernesto. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Corva, Santiago Gerardo. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Mórtola, Eduardo Carlos. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Larsen, Alejandra. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Jarvis, Donald L.. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina
Fil: Echeverria, Maria Gabriela. Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description Suid Herpesvirus 1 (SHV-1) is the etiological agent of Aujeszky’s disease (AD), which affects swine herds worldwide and causes substantial economic losses due to animal mortality and lost productivity. In order to eradicate SHV-1, vaccination programs using viruses lacking the gene encoding glycoprotein E (gE) are ongoing in several countries. These eradication programs have generated a currently unmet demand for affordable and sensitive tests that can detect SHV-1 infection, yet distinguish between infected and vaccinated pigs. To meet this demand, we used the baculovirus-insect cell system to produce immunologically authentic full-length recombinant gE protein for use in a serum ELISA assay. As previous efforts to clone the gE gene had failed due to its extremely high GC-content (75% average), we used betaine as a PCR enhancer to facilitate amplification of the entire gE gene from the Argentinian CL15 strain of SHV-1. The cloned gE gene was expressed at high levels in recombinant baculovirus-infected insect cells and reacted strongly with sera from SHV-1 infected pigs. We used the recombinant gE protein to develop a local indirect ELISA test with sensitivity and specificity comparable to currently available commercial tests. Thus, recombinant gE produced in baculovirus-infected insect cells is a viable source of antigen for the detection of SHV-1 in ELISA tests. We also provide evidence supporting a potential application of this recombinant form of gE as a SHV-1 subunit vaccine.
publishDate 2013
dc.date.none.fl_str_mv 2013-07
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/11596
Serena, Maria Soledad; Geisler, Christoph; Metz, German Ernesto; Corva, Santiago Gerardo; Mórtola, Eduardo Carlos; et al.; Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs; Elsevier; Protein Expression And Purification; 90; 1; 7-2013; 1-8
1046-5928
url http://hdl.handle.net/11336/11596
identifier_str_mv Serena, Maria Soledad; Geisler, Christoph; Metz, German Ernesto; Corva, Santiago Gerardo; Mórtola, Eduardo Carlos; et al.; Expression and purification of Suid Herpesvirus-1 glycoprotein E in the baculovirus system and its use to diagnose Aujeszky’s disease in infected pigs; Elsevier; Protein Expression And Purification; 90; 1; 7-2013; 1-8
1046-5928
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2013.04.008
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1046592813000752
info:eu-repo/semantics/altIdentifier/url/https://pubmed.ncbi.nlm.nih.gov/23631926/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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