Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.

Autores
Chimeno Zoth, Silvina Andrea; Gomez, Evangelina Raquel; Carballeda, Juan Manuel; Carrillo, Elisa Cristina; Berinstein, Analía
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV) constitutes, together with the fusion glycoprotein, the main surface antigen of this avian pathogen, which causes a highly contagious disease, relevant economically worldwide. The purpose of this work was to obtain the HN glycoprotein as a soluble antigen in culture supernatants of recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells and to evaluate its application to the development of a recombinant enzyme-linked immunosorbent assay (rELISA) for the analysis of chicken sera. A transfer vector for baculovirus containing the sequence of a melittin signal peptide was constructed and the sequence coding for HN protein without its own signal peptide was cloned. The recombinant protein was secreted and recovered easily from the culture medium of Sf9-infected cells. The recombinant protein was evaluated as antigen for ELISA coating the plates with the recovered HN using 79 positive and 142 negative samples. The Cohen kappa value resulted 0.91, indicating excellent agreement between the rELISA and the hemagglutinin inhibition tests. The rELISA was also compared with a commercial ELISA, finding high levels of agreement between both assays. The present results show that the cloning strategy developed yielded the HN protein free in the cell culture supernatant and that the recombinant protein retained its reactivity with anti-NDV HN antibodies in chicken sera.
Fil: Chimeno Zoth, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Gomez, Evangelina Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Carballeda, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Berinstein, Analía. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Materia
ELISA
HEMAGLUTININA-NEURAMINIDASA
NEWCASTLE DISEASE VIRUS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/286887
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/286887
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.Chimeno Zoth, Silvina AndreaGomez, Evangelina RaquelCarballeda, Juan ManuelCarrillo, Elisa CristinaBerinstein, AnalíaELISAHEMAGLUTININA-NEURAMINIDASANEWCASTLE DISEASE VIRUShttps://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4The hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV) constitutes, together with the fusion glycoprotein, the main surface antigen of this avian pathogen, which causes a highly contagious disease, relevant economically worldwide. The purpose of this work was to obtain the HN glycoprotein as a soluble antigen in culture supernatants of recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells and to evaluate its application to the development of a recombinant enzyme-linked immunosorbent assay (rELISA) for the analysis of chicken sera. A transfer vector for baculovirus containing the sequence of a melittin signal peptide was constructed and the sequence coding for HN protein without its own signal peptide was cloned. The recombinant protein was secreted and recovered easily from the culture medium of Sf9-infected cells. The recombinant protein was evaluated as antigen for ELISA coating the plates with the recovered HN using 79 positive and 142 negative samples. The Cohen kappa value resulted 0.91, indicating excellent agreement between the rELISA and the hemagglutinin inhibition tests. The rELISA was also compared with a commercial ELISA, finding high levels of agreement between both assays. The present results show that the cloning strategy developed yielded the HN protein free in the cell culture supernatant and that the recombinant protein retained its reactivity with anti-NDV HN antibodies in chicken sera.Fil: Chimeno Zoth, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Gomez, Evangelina Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Carballeda, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaFil: Berinstein, Analía. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; ArgentinaAmerican Association of Veterinary Laboratory Diagnosticians2011-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/286887Chimeno Zoth, Silvina Andrea; Gomez, Evangelina Raquel; Carballeda, Juan Manuel; Carrillo, Elisa Cristina; Berinstein, Analía; Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.; American Association of Veterinary Laboratory Diagnosticians; Journal of Veterinary Diagnostic Investigation; 23; 4-2011; 519-5231040-6387CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://journals.sagepub.com/doi/10.1177/1040638711404153info:eu-repo/semantics/altIdentifier/doi/10.1177/1040638711404153info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-06-17T09:38:42Zoai:ri.conicet.gov.ar:11336/286887instacron: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:34982026-06-17 09:38:42.398CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
title Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
spellingShingle Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
Chimeno Zoth, Silvina Andrea
ELISA
HEMAGLUTININA-NEURAMINIDASA
NEWCASTLE DISEASE VIRUS
title_short Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
title_full Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
title_fullStr Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
title_full_unstemmed Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
title_sort Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.
dc.creator.none.fl_str_mv Chimeno Zoth, Silvina Andrea
Gomez, Evangelina Raquel
Carballeda, Juan Manuel
Carrillo, Elisa Cristina
Berinstein, Analía
author Chimeno Zoth, Silvina Andrea
author_facet Chimeno Zoth, Silvina Andrea
Gomez, Evangelina Raquel
Carballeda, Juan Manuel
Carrillo, Elisa Cristina
Berinstein, Analía
author_role author
author2 Gomez, Evangelina Raquel
Carballeda, Juan Manuel
Carrillo, Elisa Cristina
Berinstein, Analía
author2_role author
author
author
author
dc.subject.none.fl_str_mv ELISA
HEMAGLUTININA-NEURAMINIDASA
NEWCASTLE DISEASE VIRUS
topic ELISA
HEMAGLUTININA-NEURAMINIDASA
NEWCASTLE DISEASE VIRUS
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv The hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV) constitutes, together with the fusion glycoprotein, the main surface antigen of this avian pathogen, which causes a highly contagious disease, relevant economically worldwide. The purpose of this work was to obtain the HN glycoprotein as a soluble antigen in culture supernatants of recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells and to evaluate its application to the development of a recombinant enzyme-linked immunosorbent assay (rELISA) for the analysis of chicken sera. A transfer vector for baculovirus containing the sequence of a melittin signal peptide was constructed and the sequence coding for HN protein without its own signal peptide was cloned. The recombinant protein was secreted and recovered easily from the culture medium of Sf9-infected cells. The recombinant protein was evaluated as antigen for ELISA coating the plates with the recovered HN using 79 positive and 142 negative samples. The Cohen kappa value resulted 0.91, indicating excellent agreement between the rELISA and the hemagglutinin inhibition tests. The rELISA was also compared with a commercial ELISA, finding high levels of agreement between both assays. The present results show that the cloning strategy developed yielded the HN protein free in the cell culture supernatant and that the recombinant protein retained its reactivity with anti-NDV HN antibodies in chicken sera.
Fil: Chimeno Zoth, Silvina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Gomez, Evangelina Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Carballeda, Juan Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Carrillo, Elisa Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
Fil: Berinstein, Analía. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
description The hemagglutinin-neuraminidase (HN) glycoprotein of Newcastle disease virus (NDV) constitutes, together with the fusion glycoprotein, the main surface antigen of this avian pathogen, which causes a highly contagious disease, relevant economically worldwide. The purpose of this work was to obtain the HN glycoprotein as a soluble antigen in culture supernatants of recombinant baculovirus-infected Spodoptera frugiperda (Sf9) cells and to evaluate its application to the development of a recombinant enzyme-linked immunosorbent assay (rELISA) for the analysis of chicken sera. A transfer vector for baculovirus containing the sequence of a melittin signal peptide was constructed and the sequence coding for HN protein without its own signal peptide was cloned. The recombinant protein was secreted and recovered easily from the culture medium of Sf9-infected cells. The recombinant protein was evaluated as antigen for ELISA coating the plates with the recovered HN using 79 positive and 142 negative samples. The Cohen kappa value resulted 0.91, indicating excellent agreement between the rELISA and the hemagglutinin inhibition tests. The rELISA was also compared with a commercial ELISA, finding high levels of agreement between both assays. The present results show that the cloning strategy developed yielded the HN protein free in the cell culture supernatant and that the recombinant protein retained its reactivity with anti-NDV HN antibodies in chicken sera.
publishDate 2011
dc.date.none.fl_str_mv 2011-04
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/11336/286887
Chimeno Zoth, Silvina Andrea; Gomez, Evangelina Raquel; Carballeda, Juan Manuel; Carrillo, Elisa Cristina; Berinstein, Analía; Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.; American Association of Veterinary Laboratory Diagnosticians; Journal of Veterinary Diagnostic Investigation; 23; 4-2011; 519-523
1040-6387
CONICET Digital
CONICET
url http://hdl.handle.net/11336/286887
identifier_str_mv Chimeno Zoth, Silvina Andrea; Gomez, Evangelina Raquel; Carballeda, Juan Manuel; Carrillo, Elisa Cristina; Berinstein, Analía; Expression of a secreted version of the HN glycoprotein of Newcastle disease virus: its evaluation as a diagnostic reagent.; American Association of Veterinary Laboratory Diagnosticians; Journal of Veterinary Diagnostic Investigation; 23; 4-2011; 519-523
1040-6387
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://journals.sagepub.com/doi/10.1177/1040638711404153
info:eu-repo/semantics/altIdentifier/doi/10.1177/1040638711404153
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Association of Veterinary Laboratory Diagnosticians
publisher.none.fl_str_mv American Association of Veterinary Laboratory Diagnosticians
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 13.040872

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