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A new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulations

2012-01Journal article Open access
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dc.contributor.authorBasto, Afonso P.
dc.contributor.authorPiedade, João
dc.contributor.authorRamalho, Ruben
dc.contributor.authorAlves, Susana
dc.contributor.authorSoares, Helena
dc.contributor.authorCornelis, Pierre
dc.contributor.authorMartins, Carlos
dc.contributor.authorLeitão, Alexandre
dc.date.accessioned2013-02-01T15:01:14Z
dc.date.available2013-02-01T15:01:14Z
dc.date.issued2012-01
dc.description.abstractThe 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.por
dc.identifier.citationBasto 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.por
dc.identifier.issn0168-1656
dc.identifier.urihttp://hdl.handle.net/10400.21/2159
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherElsevierpor
dc.relation.publisherversionhttp://www.sciencedirect.com/science/article/pii/S0168165611006407por
dc.subjectAfrican swine fever virus/geneticspor
dc.subjectAnimalspor
dc.subjectAntigens, viral/geneticspor
dc.subjectBacterial proteins/biosynthesispor
dc.subjectBacterial proteins/geneticspor
dc.subjectBacterial proteins/immunologypor
dc.subjectBase sequencepor
dc.subjectCells, culturedpor
dc.subjectCloning, molecular/methodspor
dc.subjectDendritic cells/immunologypor
dc.subjectEscherichia coli/geneticspor
dc.subjectEscherichia coli/metabolismpor
dc.subjectGenetic vectors/geneticspor
dc.subjectLipoproteins/biosynthesispor
dc.subjectLipoproteins/geneticspor
dc.subjectLipoproteins/immunologypor
dc.subjectMacrophages/immunologypor
dc.subjectMicepor
dc.subjectMolecular sequence datapor
dc.subjectPseudomonas aeruginosa/geneticspor
dc.subjectRecombinant fusion proteins/biosynthesispor
dc.subjectRecombinant fusion proteins/geneticspor
dc.subjectRecombinant fusion proteins/immunologypor
dc.subjectTumor necrosis factor-alpha/metabolismpor
dc.titleA new cloning system based on the OprI lipoprotein for the production of recombinant bacterial cell wall-derived immunogenic formulationspor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage63por
oaire.citation.startPage50por
oaire.citation.titleJournal of Biotechnologypor
oaire.citation.volume157por
rcaap.rightsembargoedAccesspor
rcaap.typearticlepor

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