The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system

Autores
Marcipar, Iván Sergio; Olivares, María Laura; Robles, Lucia; Dekanty, Andres; Marcipar, Alberto Jaime; Silber, Ariel Mariano
Año de publicación
2004
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In the present work, we have determined the effect of expression vectors and their corresponding host bacteria on the antigenic performance of Trypanosoma cruzi P2b (TcP2bÞ full-length recombinant protein. The gene encoding the TcP2b ribosomal protein was cloned in pMAL-c2 and pET-32a vectors that allow the expression of high levels of soluble fusion proteins. A panel of 32 positive and 32 negative sera was assayed with the purified proteins expressed using pMal-c2 (TcP2b– MBP) and pET-32a (TcP2b–TRX) vectors and with MBP and TRX purified from pMAL-c2 and pET-32a vectors, respectively. The antigenic behavior of each TcP2b recombinant protein differed in the diagnostic performance in terms of DI(+) (93.7 for TcP2b–MBP vs 100% for TcP2b–TRX), in DI()) (90.5 for TcP2b–MBP vs 100% for TcP2b–TRX) and in cross-reaction with negative sera. To determine if the higher reactivity of expressed pMAL-c2 protein was due to folding during protein expression or to a steric effect related to the protein adsorption at the titration plate, the reactivity of sera against soluble proteins was assessed by ELISA inhibition assays. As each soluble protein preserved its level of reactivity, we concluded that differences in reactivity were due to intrinsic characteristics of the proteins and not to differences in patterns of adsorption to the plates.
Fil: Marcipar, Iván Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Olivares, María Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Robles, Lucia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina
Fil: Dekanty, Andres. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Marcipar, Alberto Jaime. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina
Fil: Silber, Ariel Mariano. Universidade de Sao Paulo; Brasil
Materia
Trypanosoma Cruzi
P2beta
Antigen
Diagnostic
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:11336/26821
Acceder
id CONICETDig_913c7b59ceda7861125321d22a9ff761
oai_identifier_str oai:ri.conicet.gov.ar:11336/26821
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression systemMarcipar, Iván SergioOlivares, María LauraRobles, LuciaDekanty, AndresMarcipar, Alberto JaimeSilber, Ariel MarianoTrypanosoma CruziP2betaAntigenDiagnostichttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1In the present work, we have determined the effect of expression vectors and their corresponding host bacteria on the antigenic performance of Trypanosoma cruzi P2b (TcP2bÞ full-length recombinant protein. The gene encoding the TcP2b ribosomal protein was cloned in pMAL-c2 and pET-32a vectors that allow the expression of high levels of soluble fusion proteins. A panel of 32 positive and 32 negative sera was assayed with the purified proteins expressed using pMal-c2 (TcP2b– MBP) and pET-32a (TcP2b–TRX) vectors and with MBP and TRX purified from pMAL-c2 and pET-32a vectors, respectively. The antigenic behavior of each TcP2b recombinant protein differed in the diagnostic performance in terms of DI(+) (93.7 for TcP2b–MBP vs 100% for TcP2b–TRX), in DI()) (90.5 for TcP2b–MBP vs 100% for TcP2b–TRX) and in cross-reaction with negative sera. To determine if the higher reactivity of expressed pMAL-c2 protein was due to folding during protein expression or to a steric effect related to the protein adsorption at the titration plate, the reactivity of sera against soluble proteins was assessed by ELISA inhibition assays. As each soluble protein preserved its level of reactivity, we concluded that differences in reactivity were due to intrinsic characteristics of the proteins and not to differences in patterns of adsorption to the plates.Fil: Marcipar, Iván Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Olivares, María Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Robles, Lucia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; ArgentinaFil: Dekanty, Andres. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Marcipar, Alberto Jaime. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; ArgentinaFil: Silber, Ariel Mariano. Universidade de Sao Paulo; BrasilElsevier Inc2004-01info: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/11336/26821Marcipar, Iván Sergio; Olivares, María Laura; Robles, Lucia; Dekanty, Andres; Marcipar, Alberto Jaime; et al.; The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system; Elsevier Inc; Protein Expression and Purification; 34; 1; 1-2004; 1-71046-5928CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2003.11.022info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1046592803003875info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:09:58Zoai:ri.conicet.gov.ar:11336/26821instacron: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:34982025-10-22 11:09:58.518CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
title The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
spellingShingle The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
Marcipar, Iván Sergio
Trypanosoma Cruzi
P2beta
Antigen
Diagnostic
title_short The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
title_full The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
title_fullStr The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
title_full_unstemmed The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
title_sort The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system
dc.creator.none.fl_str_mv Marcipar, Iván Sergio
Olivares, María Laura
Robles, Lucia
Dekanty, Andres
Marcipar, Alberto Jaime
Silber, Ariel Mariano
author Marcipar, Iván Sergio
author_facet Marcipar, Iván Sergio
Olivares, María Laura
Robles, Lucia
Dekanty, Andres
Marcipar, Alberto Jaime
Silber, Ariel Mariano
author_role author
author2 Olivares, María Laura
Robles, Lucia
Dekanty, Andres
Marcipar, Alberto Jaime
Silber, Ariel Mariano
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Trypanosoma Cruzi
P2beta
Antigen
Diagnostic
topic Trypanosoma Cruzi
P2beta
Antigen
Diagnostic
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv In the present work, we have determined the effect of expression vectors and their corresponding host bacteria on the antigenic performance of Trypanosoma cruzi P2b (TcP2bÞ full-length recombinant protein. The gene encoding the TcP2b ribosomal protein was cloned in pMAL-c2 and pET-32a vectors that allow the expression of high levels of soluble fusion proteins. A panel of 32 positive and 32 negative sera was assayed with the purified proteins expressed using pMal-c2 (TcP2b– MBP) and pET-32a (TcP2b–TRX) vectors and with MBP and TRX purified from pMAL-c2 and pET-32a vectors, respectively. The antigenic behavior of each TcP2b recombinant protein differed in the diagnostic performance in terms of DI(+) (93.7 for TcP2b–MBP vs 100% for TcP2b–TRX), in DI()) (90.5 for TcP2b–MBP vs 100% for TcP2b–TRX) and in cross-reaction with negative sera. To determine if the higher reactivity of expressed pMAL-c2 protein was due to folding during protein expression or to a steric effect related to the protein adsorption at the titration plate, the reactivity of sera against soluble proteins was assessed by ELISA inhibition assays. As each soluble protein preserved its level of reactivity, we concluded that differences in reactivity were due to intrinsic characteristics of the proteins and not to differences in patterns of adsorption to the plates.
Fil: Marcipar, Iván Sergio. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Olivares, María Laura. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Robles, Lucia. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina
Fil: Dekanty, Andres. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Marcipar, Alberto Jaime. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Instituto de Tecnología Biológica; Argentina
Fil: Silber, Ariel Mariano. Universidade de Sao Paulo; Brasil
description In the present work, we have determined the effect of expression vectors and their corresponding host bacteria on the antigenic performance of Trypanosoma cruzi P2b (TcP2bÞ full-length recombinant protein. The gene encoding the TcP2b ribosomal protein was cloned in pMAL-c2 and pET-32a vectors that allow the expression of high levels of soluble fusion proteins. A panel of 32 positive and 32 negative sera was assayed with the purified proteins expressed using pMal-c2 (TcP2b– MBP) and pET-32a (TcP2b–TRX) vectors and with MBP and TRX purified from pMAL-c2 and pET-32a vectors, respectively. The antigenic behavior of each TcP2b recombinant protein differed in the diagnostic performance in terms of DI(+) (93.7 for TcP2b–MBP vs 100% for TcP2b–TRX), in DI()) (90.5 for TcP2b–MBP vs 100% for TcP2b–TRX) and in cross-reaction with negative sera. To determine if the higher reactivity of expressed pMAL-c2 protein was due to folding during protein expression or to a steric effect related to the protein adsorption at the titration plate, the reactivity of sera against soluble proteins was assessed by ELISA inhibition assays. As each soluble protein preserved its level of reactivity, we concluded that differences in reactivity were due to intrinsic characteristics of the proteins and not to differences in patterns of adsorption to the plates.
publishDate 2004
dc.date.none.fl_str_mv 2004-01
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/11336/26821
Marcipar, Iván Sergio; Olivares, María Laura; Robles, Lucia; Dekanty, Andres; Marcipar, Alberto Jaime; et al.; The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system; Elsevier Inc; Protein Expression and Purification; 34; 1; 1-2004; 1-7
1046-5928
CONICET Digital
CONICET
url http://hdl.handle.net/11336/26821
identifier_str_mv Marcipar, Iván Sergio; Olivares, María Laura; Robles, Lucia; Dekanty, Andres; Marcipar, Alberto Jaime; et al.; The diagnostic performance of recombinant Trypanosoma cruzi ribosomal P2β protein is influenced by its expression system; Elsevier Inc; Protein Expression and Purification; 34; 1; 1-2004; 1-7
1046-5928
CONICET Digital
CONICET
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.2003.11.022
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1046592803003875
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 Inc
publisher.none.fl_str_mv Elsevier Inc
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 12.982451

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