A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.21/2159 |
Resumo: | The conjugation of antigens with ligands of pattern recognition receptors (PRR) is emerging as a promising strategy for the modulation of specific immunity. Here, we describe a new Escherichia coli system for the cloning and expression of heterologous antigens in fusion with the OprI lipoprotein, a TLR ligand from the Pseudomonas aeruginosa outer membrane (OM). Analysis of the OprI expressed by this system reveals a triacylated lipid moiety mainly composed by palmitic acid residues. By offering a tight regulation of expression and allowing for antigen purification by metal affinity chromatography, the new system circumvents the major drawbacks of former versions. In addition, the anchoring of OprI to the OM of the host cell is further explored for the production of novel recombinant bacterial cell wall-derived formulations (OM fragments and OM vesicles) with distinct potential for PRR activation. As an example, the African swine fever virus ORF A104R was cloned and the recombinant antigen was obtained in the three formulations. Overall, our results validate a new system suitable for the production of immunogenic formulations that can be used for the development of experimental vaccines and for studies on the modulation of acquired immunity. |
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A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulationsAfrican swine fever virus/geneticsAnimalsAntigens, viral/geneticsBacterial proteins/biosynthesisBacterial proteins/geneticsBacterial proteins/immunologyBase sequenceCells, culturedCloning, molecular/methodsDendritic cells/immunologyEscherichia coli/geneticsEscherichia coli/metabolismGenetic vectors/geneticsLipoproteins/biosynthesisLipoproteins/geneticsLipoproteins/immunologyMacrophages/immunologyMiceMolecular sequence dataPseudomonas aeruginosa/geneticsRecombinant fusion proteins/biosynthesisRecombinant fusion proteins/geneticsRecombinant fusion proteins/immunologyTumor necrosis factor-alpha/metabolismThe conjugation of antigens with ligands of pattern recognition receptors (PRR) is emerging as a promising strategy for the modulation of specific immunity. Here, we describe a new Escherichia coli system for the cloning and expression of heterologous antigens in fusion with the OprI lipoprotein, a TLR ligand from the Pseudomonas aeruginosa outer membrane (OM). Analysis of the OprI expressed by this system reveals a triacylated lipid moiety mainly composed by palmitic acid residues. By offering a tight regulation of expression and allowing for antigen purification by metal affinity chromatography, the new system circumvents the major drawbacks of former versions. In addition, the anchoring of OprI to the OM of the host cell is further explored for the production of novel recombinant bacterial cell wall-derived formulations (OM fragments and OM vesicles) with distinct potential for PRR activation. As an example, the African swine fever virus ORF A104R was cloned and the recombinant antigen was obtained in the three formulations. Overall, our results validate a new system suitable for the production of immunogenic formulations that can be used for the development of experimental vaccines and for studies on the modulation of acquired immunity.ElsevierRCIPLBasto, Afonso P.Piedade, JoãoRamalho, RubenAlves, SusanaSoares, HelenaCornelis, PierreMartins, CarlosLeitão, Alexandre2013-02-01T15:01:14Z2012-012012-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.21/2159engBasto AP, Piedade J, Ramalho R, Alves S, Soares H, Cornelis P, et al. A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations. J Biotechnol. 2012;157(1):50-63.0168-1656info:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-08-03T09:40:59Zoai:repositorio.ipl.pt:10400.21/2159Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:11:58.823789Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations |
title |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations |
spellingShingle |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations Basto, Afonso P. African swine fever virus/genetics Animals Antigens, viral/genetics Bacterial proteins/biosynthesis Bacterial proteins/genetics Bacterial proteins/immunology Base sequence Cells, cultured Cloning, molecular/methods Dendritic cells/immunology Escherichia coli/genetics Escherichia coli/metabolism Genetic vectors/genetics Lipoproteins/biosynthesis Lipoproteins/genetics Lipoproteins/immunology Macrophages/immunology Mice Molecular sequence data Pseudomonas aeruginosa/genetics Recombinant fusion proteins/biosynthesis Recombinant fusion proteins/genetics Recombinant fusion proteins/immunology Tumor necrosis factor-alpha/metabolism |
title_short |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations |
title_full |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations |
title_fullStr |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations |
title_full_unstemmed |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations |
title_sort |
A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations |
author |
Basto, Afonso P. |
author_facet |
Basto, Afonso P. Piedade, João Ramalho, Ruben Alves, Susana Soares, Helena Cornelis, Pierre Martins, Carlos Leitão, Alexandre |
author_role |
author |
author2 |
Piedade, João Ramalho, Ruben Alves, Susana Soares, Helena Cornelis, Pierre Martins, Carlos Leitão, Alexandre |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
RCIPL |
dc.contributor.author.fl_str_mv |
Basto, Afonso P. Piedade, João Ramalho, Ruben Alves, Susana Soares, Helena Cornelis, Pierre Martins, Carlos Leitão, Alexandre |
dc.subject.por.fl_str_mv |
African swine fever virus/genetics Animals Antigens, viral/genetics Bacterial proteins/biosynthesis Bacterial proteins/genetics Bacterial proteins/immunology Base sequence Cells, cultured Cloning, molecular/methods Dendritic cells/immunology Escherichia coli/genetics Escherichia coli/metabolism Genetic vectors/genetics Lipoproteins/biosynthesis Lipoproteins/genetics Lipoproteins/immunology Macrophages/immunology Mice Molecular sequence data Pseudomonas aeruginosa/genetics Recombinant fusion proteins/biosynthesis Recombinant fusion proteins/genetics Recombinant fusion proteins/immunology Tumor necrosis factor-alpha/metabolism |
topic |
African swine fever virus/genetics Animals Antigens, viral/genetics Bacterial proteins/biosynthesis Bacterial proteins/genetics Bacterial proteins/immunology Base sequence Cells, cultured Cloning, molecular/methods Dendritic cells/immunology Escherichia coli/genetics Escherichia coli/metabolism Genetic vectors/genetics Lipoproteins/biosynthesis Lipoproteins/genetics Lipoproteins/immunology Macrophages/immunology Mice Molecular sequence data Pseudomonas aeruginosa/genetics Recombinant fusion proteins/biosynthesis Recombinant fusion proteins/genetics Recombinant fusion proteins/immunology Tumor necrosis factor-alpha/metabolism |
description |
The conjugation of antigens with ligands of pattern recognition receptors (PRR) is emerging as a promising strategy for the modulation of specific immunity. Here, we describe a new Escherichia coli system for the cloning and expression of heterologous antigens in fusion with the OprI lipoprotein, a TLR ligand from the Pseudomonas aeruginosa outer membrane (OM). Analysis of the OprI expressed by this system reveals a triacylated lipid moiety mainly composed by palmitic acid residues. By offering a tight regulation of expression and allowing for antigen purification by metal affinity chromatography, the new system circumvents the major drawbacks of former versions. In addition, the anchoring of OprI to the OM of the host cell is further explored for the production of novel recombinant bacterial cell wall-derived formulations (OM fragments and OM vesicles) with distinct potential for PRR activation. As an example, the African swine fever virus ORF A104R was cloned and the recombinant antigen was obtained in the three formulations. Overall, our results validate a new system suitable for the production of immunogenic formulations that can be used for the development of experimental vaccines and for studies on the modulation of acquired immunity. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-01 2012-01-01T00:00:00Z 2013-02-01T15:01:14Z |
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://hdl.handle.net/10400.21/2159 |
url |
http://hdl.handle.net/10400.21/2159 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Basto AP, Piedade J, Ramalho R, Alves S, Soares H, Cornelis P, et al. A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations. J Biotechnol. 2012;157(1):50-63. 0168-1656 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799133374778966016 |