Optimization and scale-up production of Zika virus deltaNS1 in Escherichia coli: application of Response Surface Methodology

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dc.contributorLEDV - Laboratório de Desenvolvimento de Vacinaspt_BR
dc.contributor.authorKanno, Alex Issamupt_BR
dc.contributor.authorLeite, Luciana Cezar de Cerqueirapt_BR
dc.contributor.authorPereira, Lennon Ramospt_BR
dc.contributor.authorJesus, Mônica Josiane Rodrigues dept_BR
dc.contributor.authorAndreata-Santos, Robertpt_BR
dc.contributor.authorAlves, Rúbens Prince dos Santospt_BR
dc.contributor.authorDurigon, Edison Luizpt_BR
dc.contributor.authorGonçalves, Viviane Maimonipt_BR
dc.date.accessioned2020-07-09T21:25:59Z-
dc.date.available2020-07-09T21:25:59Z-
dc.date.issued2019pt_BR
dc.identifier.citationKanno AI, Leite LCC, Pereira LR, Jesus MJR, Andreata-Santos R, Alves RPS, et al. Optimization and scale-up production of Zika virus deltaNS1 in Escherichia coli: application of Response Surface Methodology. AMB Express. 2019 Dec;10:1. doi:10.1186/s13568-019-0926-y.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/2899-
dc.description.abstractDiagnosing Zika virus (ZIKV) infections has been challenging due to the cross-reactivity of induced antibodies with other flavivirus. The concomitant occurrence of ZIKV and Dengue virus (DENV) in endemic regions requires diagnostic tools with the ability to distinguish these two viral infections. Recent studies demonstrated that immunoassays using the C-terminal fragment of ZIKV NS1 antigen (DeltaNS1) can be used to discriminate ZIKV from DENV infections. In order to be used in serological tests, the expression/solubility of DeltaNS1 and growth of recombinant E. coli strain were optimized by Response Surface Methodology. Temperature, time and IPTG concentration were evaluated. According to the model, the best condition determined in small scale cultures was 21 °C for 20 h with 0.7 mM of IPTG, which predicted 7.5 g/L of biomass and 962 mg/L of DeltaNS1. These conditions were validated and used in a 6-L batch in the bioreactor, which produced 6.4 g/L of biomass and 500 mg/L of DeltaNS1 in 12 h of induction. The serological ELISA test performed with purified DeltaNS1 showed low cross-reactivity with antibodies from DENV-infected human subjects. Denaturation of DeltaNS1 decreased the detection of anti-ZIKV antibodies, thus indicating the contribution of conformational epitopes and confirming the importance of properly folded DeltaNS1 for the specificity of the serological analyses. Obtaining high yields of soluble DeltaNS1 supports the viability of an effective serologic diagnostic test capable of differentiating ZIKV from other flavivirus infections.pt_BR
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)pt_BR
dc.format.extent1pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofAMB Expresspt_BR
dc.rightsOpen Accesspt_BR
dc.titleOptimization and scale-up production of Zika virus deltaNS1 in Escherichia coli: application of Response Surface Methodologypt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1186/s13568-019-0926-ypt_BR
dc.identifier.urlhttps://doi.org/10.1186/s13568-019-0926-ypt_BR
dc.contributor.externalUniversidade de São Paulo (USP)¦¦Brasilpt_BR
dc.identifier.citationvolume10pt_BR
dc.subject.keywordZika NS1pt_BR
dc.subject.keywordSerological diagnosispt_BR
dc.subject.keywordE. colipt_BR
dc.subject.keywordHeterologous protein productionpt_BR
dc.subject.keywordSoluble expressionpt_BR
dc.subject.keywordResponse Surface Methodologypt_BR
dc.relation.ispartofabbreviatedAMB Expresspt_BR
dc.identifier.citationabntv. 10, 1, dec. 2019pt_BR
dc.identifier.citationvancouver2019 Dec;10:1pt_BR
dc.contributor.butantanKanno, Alex Issamu|:Pesquisador|:LEDV - Laboratório de Desenvolvimento de Vacinas|:PrimeiroAutorpt_BR
dc.contributor.butantanLeite, Luciana Cezar de Cerqueira|:Pesquisador|:LEDV - Laboratório de Desenvolvimento de Vacinas|:pt_BR
dc.contributor.butantanGonçalves, Viviane Maimoni|:Pesquisador|:LEDV - Laboratório de Desenvolvimento de Vacinas|:Autor de correspondênciapt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2012/23124-4pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2016/20045-7pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2014/17595-0pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.openairetypeArticle-
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item.languageiso639-1English-
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