Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)

Detalhes bibliográficos
Autor(a) principal: Farias, Thais H.v.
Data de Publicação: 2018
Outros Autores: Silva, Ketherson R., Mariguela, Viviane C., Montassier, HÉlio J., Pilarski, Fabiana
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1590/0001-3765201820170438
http://hdl.handle.net/11449/158090
Resumo: ABSTRACT The pacu is one of the most important species for Brazilian fish farming and is considered emerging in the global aquaculture. Despite its importance, no effective tool for evaluation of the adaptive immune response of this species has been developed. Therefore, this study aimed the development and standardization of indirect ELISA for the measurement of pacu antigen-specific antibodies using polyclonal rabbit anti-pacu IgM used as detector antibody. For this purpose was isolated and purificated of pacu IgM using mannose-binding protein affinity chromatography and produced specific polyclonal antibodies against heavy and light chains pacu IgM, that showed a molecular weight of 72 kDa and 26 kDa, respectively. Polyclonal antibodies obtained demonstrated specificity with heavy and light Ig chains of pacu serum in western blotting. These polyclonal antibodies allowed the development of an indirect ELISA assay of high sensitivity and specificity for the detection and quantification of pacu IgM antibodies immunized with bovine IgG. In conclusion, this approach has great potential to improve the monitoring of vaccine-induced immune responses and help develop immunodiagnostic and epidemiological studies of infectious diseases in pacu systems.
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spelling Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)ELISAImmunoglobulinPacu`s immune systemPolyclonal antiseraABSTRACT The pacu is one of the most important species for Brazilian fish farming and is considered emerging in the global aquaculture. Despite its importance, no effective tool for evaluation of the adaptive immune response of this species has been developed. Therefore, this study aimed the development and standardization of indirect ELISA for the measurement of pacu antigen-specific antibodies using polyclonal rabbit anti-pacu IgM used as detector antibody. For this purpose was isolated and purificated of pacu IgM using mannose-binding protein affinity chromatography and produced specific polyclonal antibodies against heavy and light chains pacu IgM, that showed a molecular weight of 72 kDa and 26 kDa, respectively. Polyclonal antibodies obtained demonstrated specificity with heavy and light Ig chains of pacu serum in western blotting. These polyclonal antibodies allowed the development of an indirect ELISA assay of high sensitivity and specificity for the detection and quantification of pacu IgM antibodies immunized with bovine IgG. In conclusion, this approach has great potential to improve the monitoring of vaccine-induced immune responses and help develop immunodiagnostic and epidemiological studies of infectious diseases in pacu systems.Universidade Estadual Paulista Centro de Aquicultura Laboratório de Patologia de Organismos AquáticosCentro Universitário Barão de MauáUniversidade Estadual Paulista, Campus Jaboticabal Faculdade de Ciências Agrárias e Veterinárias Departamento de Patologia VeterináriaUniversidade Estadual Paulista Centro de Aquicultura Laboratório de Patologia de Organismos AquáticosUniversidade Estadual Paulista, Campus Jaboticabal Faculdade de Ciências Agrárias e Veterinárias Departamento de Patologia VeterináriaAcademia Brasileira de CiênciasUniversidade Estadual Paulista (Unesp)Centro Universitário Barão de MauáFarias, Thais H.v.Silva, Ketherson R.Mariguela, Viviane C.Montassier, HÉlio J.Pilarski, Fabiana2018-11-12T17:28:16Z2018-11-12T17:28:16Z2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article0-0application/pdfhttp://dx.doi.org/10.1590/0001-3765201820170438Anais da Academia Brasileira de Ciências. Academia Brasileira de Ciências, n. ahead, p. 0-0, 2018.0001-3765http://hdl.handle.net/11449/15809010.1590/0001-3765201820170438S0001-37652018005011105S0001-37652018005011105.pdfSciELOreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengAnais da Academia Brasileira de Ciências0,418info:eu-repo/semantics/openAccess2024-06-07T13:02:57Zoai:repositorio.unesp.br:11449/158090Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T21:40:13.872999Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
title Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
spellingShingle Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
Farias, Thais H.v.
ELISA
Immunoglobulin
Pacu`s immune system
Polyclonal antisera
title_short Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
title_full Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
title_fullStr Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
title_full_unstemmed Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
title_sort Development of an indirect ELISA assay to evaluation of the adaptive immune response of pacu (Piaractus mesopotamicus)
author Farias, Thais H.v.
author_facet Farias, Thais H.v.
Silva, Ketherson R.
Mariguela, Viviane C.
Montassier, HÉlio J.
Pilarski, Fabiana
author_role author
author2 Silva, Ketherson R.
Mariguela, Viviane C.
Montassier, HÉlio J.
Pilarski, Fabiana
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
Centro Universitário Barão de Mauá
dc.contributor.author.fl_str_mv Farias, Thais H.v.
Silva, Ketherson R.
Mariguela, Viviane C.
Montassier, HÉlio J.
Pilarski, Fabiana
dc.subject.por.fl_str_mv ELISA
Immunoglobulin
Pacu`s immune system
Polyclonal antisera
topic ELISA
Immunoglobulin
Pacu`s immune system
Polyclonal antisera
description ABSTRACT The pacu is one of the most important species for Brazilian fish farming and is considered emerging in the global aquaculture. Despite its importance, no effective tool for evaluation of the adaptive immune response of this species has been developed. Therefore, this study aimed the development and standardization of indirect ELISA for the measurement of pacu antigen-specific antibodies using polyclonal rabbit anti-pacu IgM used as detector antibody. For this purpose was isolated and purificated of pacu IgM using mannose-binding protein affinity chromatography and produced specific polyclonal antibodies against heavy and light chains pacu IgM, that showed a molecular weight of 72 kDa and 26 kDa, respectively. Polyclonal antibodies obtained demonstrated specificity with heavy and light Ig chains of pacu serum in western blotting. These polyclonal antibodies allowed the development of an indirect ELISA assay of high sensitivity and specificity for the detection and quantification of pacu IgM antibodies immunized with bovine IgG. In conclusion, this approach has great potential to improve the monitoring of vaccine-induced immune responses and help develop immunodiagnostic and epidemiological studies of infectious diseases in pacu systems.
publishDate 2018
dc.date.none.fl_str_mv 2018-11-12T17:28:16Z
2018-11-12T17:28:16Z
2018
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1590/0001-3765201820170438
Anais da Academia Brasileira de Ciências. Academia Brasileira de Ciências, n. ahead, p. 0-0, 2018.
0001-3765
http://hdl.handle.net/11449/158090
10.1590/0001-3765201820170438
S0001-37652018005011105
S0001-37652018005011105.pdf
url http://dx.doi.org/10.1590/0001-3765201820170438
http://hdl.handle.net/11449/158090
identifier_str_mv Anais da Academia Brasileira de Ciências. Academia Brasileira de Ciências, n. ahead, p. 0-0, 2018.
0001-3765
10.1590/0001-3765201820170438
S0001-37652018005011105
S0001-37652018005011105.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Anais da Academia Brasileira de Ciências
0,418
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 0-0
application/pdf
dc.publisher.none.fl_str_mv Academia Brasileira de Ciências
publisher.none.fl_str_mv Academia Brasileira de Ciências
dc.source.none.fl_str_mv SciELO
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
_version_ 1808129345418428416

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