Optimization of expression and purification of schistosoma mansoni antigens in fusion with Rhizavidin

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dc.contributorLaboratório de Desenvolvimento de Vacinaspt_BR
dc.contributor.authorBarbosa, Mayra Mara Ferraript_BR
dc.contributor.authorKanno, Alex Issamupt_BR
dc.contributor.authorPancakova, Violetapt_BR
dc.contributor.authorGonçalves, Viviane Maimonipt_BR
dc.contributor.authorMalley, Richardpt_BR
dc.contributor.authorFaria, Leonardo P.pt_BR
dc.contributor.authorLeite, Luciana Cezar de Cerqueirapt_BR
dc.date.accessioned2021-06-25T20:22:06Z-
dc.date.available2021-06-25T20:22:06Z-
dc.date.issued2021pt_BR
dc.identifier.citationBarbosa MMF, Kanno AI, Pancakova V, Gonçalves VM, Malley R, Faria LP., et al. Optimization of expression and purification of schistosoma mansoni antigens in fusion with Rhizavidin. Mol. Biotechnol.. 2021 June;63:983–991. doi:10.1007/s12033-021-00355-2.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/3868-
dc.description.abstractSchistosomiasis causes significant morbidity and mortality. Vaccine efforts to date indicate the need to increase the immunogenicity of Schistosoma antigens. The multiple antigen-presenting system, whereby proteins are genetically fused to rhizavidin and affinity linked to biotinylated templates, enables the generation of robust immune responses. The objective of this work was to express and purify the S. mansoni antigens, SmTSP-2 and SmCD59.2, in fusion with rhizavidin. The fusion with rhizavidin greatly decreased the expression level of rSmTSP-2, but not rSmCD59.2, and both were expressed in the insoluble fraction, requiring optimization of culture conditions. Evaluation of different E. coli strains and media showed that BL21-DE3 cultured in Terrific Broth provided the highest expression levels of both proteins. Investigation of a range of time and temperature of induction showed that E. coli strains expressing rRzv:SmTSP-2 and rRzv:SmCD59.2 showed the highest protein production at 23 °C for 15 h. Recombinant proteins were purified by a single step of affinity chromatography allowing isolation of these proteins in high concentration and purity. The optimization process increased final soluble protein yield of rRzv:SmTSP-2 by fourfold and rRzv:SmCD59.2 by tenfold, providing ~ 20 mg/L of each protein. Optimized fusion protein production will allow antigen use in biotin–rhizavidin affinity platforms.pt_BR
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)pt_BR
dc.description.sponsorshipFundação Butantanpt_BR
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)pt_BR
dc.format.extent983–991pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofMolecular Biotechnologypt_BR
dc.rightsRestricted accesspt_BR
dc.titleOptimization of expression and purification of schistosoma mansoni antigens in fusion with Rhizavidinpt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1007/s12033-021-00355-2pt_BR
dc.identifier.urlhttps://doi.org/10.1007/s12033-021-00355-2pt_BR
dc.contributor.externalUniversidade de São Paulo (USP)pt_BR
dc.contributor.externalBoston Children's Hospitalpt_BR
dc.contributor.externalFundação Oswaldo Cruz (Fiocruz)pt_BR
dc.contributor.externalUniversité Claude Bernard Lyon 1pt_BR
dc.identifier.citationvolume63pt_BR
dc.subject.keywordProtein expressionpt_BR
dc.subject.keywordProtein purificationpt_BR
dc.subject.keywordFusion proteinspt_BR
dc.subject.keywordRhizavidin–biotin systempt_BR
dc.subject.keywordSchistosoma proteinspt_BR
dc.relation.ispartofabbreviatedMol. Biotechnol.pt_BR
dc.identifier.citationabntv. 63, p. 983–991, jun. 2021pt_BR
dc.identifier.citationvancouver2021 June;63:983–991pt_BR
dc.contributor.butantanBarbosa, Mayra Mara Ferrari|:Aluno|:Laboratório de Desenvolvimento de Vacinas|:PrimeiroAutorpt_BR
dc.contributor.butantanKanno, Alex Issamu|:Aluno|:Laboratório de Desenvolvimento de Vacinaspt_BR
dc.contributor.butantanGonçalves, Viviane Maimoni|:Pesquisador|:Laboratório de Desenvolvimento de Vacinaspt_BR
dc.contributor.butantanLeite, Luciana Cezar de Cerqueira|:Pesquisador|:Laboratório de Desenvolvimento de Vacinas|:Autor de correspondênciapt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2017/24632-6pt_BR
dc.sponsorship.butantanFundação Butantan¦¦pt_BR
dc.sponsorship.butantanCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)¦¦pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.languageiso639-1English-
item.fulltextCom Texto completo-
item.openairetypeArticle-
item.grantfulltextembargo_29990101-
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