DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein

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dc.contributorLab. Biotecnologia Viralpt_BR
dc.contributor.authorDecarli, Monize Caiadopt_BR
dc.contributor.authordos Santos, Diogo Perespt_BR
dc.contributor.authorAstray, Renato Mancinipt_BR
dc.contributor.authorVentini, Daniella Cristinapt_BR
dc.contributor.authorJorge, Soraia Attie Calilpt_BR
dc.contributor.authorCorreia, Daniela Matildept_BR
dc.contributor.authorda Silva, Juliana de Sapt_BR
dc.contributor.authorRocca, Mayra Pereirapt_BR
dc.contributor.authorLangoni, Heliopt_BR
dc.contributor.authorMenozzi, Benedito Donizetept_BR
dc.contributor.authorPereira, Carlos Augustopt_BR
dc.contributor.authorTorres Suazo, Claudio Albertopt_BR
dc.date.accessioned2020-07-09T21:20:39Z-
dc.date.available2020-07-09T21:20:39Z-
dc.date.issued2018pt_BR
dc.identifier.citationDecarli MC, dos Santos DP, Astray RM, Ventini DC, Jorge SAC, Correia DM, et al. DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein. Appl. Microbiol. Biotechnol.. 2018 Jun;102(11):4773-83. doi:10.1007/s00253-018-8962-0.pt_BR
dc.identifier.urihttps://repositorio.butantan.gov.br/handle/butantan/2508-
dc.description.abstractThe transmembrane rabies virus glycoprotein (RVGP) is the main antigen of vaccine formulations used around the world to prevent rabies, the most lethal preventable infectious disease known. The objective of this work was to evaluate the potential of a bioreactor using wave-induced agitation in the initial steps of scaling up the rRVGP production process by a Drosophila melanogaster S2 cell line to produce rRVGP in sufficient quantities for immunization and characterization studies. Taking advantage of some remarkable features recognized in Drosophila S2 cells for scaling the culture process, a robust recombinant lineage (S2MtRVGPH-His) engineered by our group for the expression of rRVGP using a copper-inducible promoter was used in the bioreactor cultures. The WAVE Bioreactor was chosen because it represents an innovative approach to the cultivation of animal cells using single-use technology. For that purpose, we firstly established a procedure for culturing the S2MtRVGPH-His lineage in 100 mL Schott flasks. Using an inoculum of 5 x 10(5) cells/mL in culture medium (Sf900-III) induced with solution of CuSO4 (0.7 mM) and a convenient pH range (6.2-7.0), optimal parameter values such as time of induction (72 h) and temperature (28 degrees C) to increase rRVGP production could be defined. This procedure was reproduced in culture experiments conducted in a WAVE Bioreactor (TM) 2/10 using a 2 L Cellbag. The results in Schott flasks and inWAVE Bioreactor (TM) were very similar, yielding a maximum titer of rRVGP above of 1 mg.L-1. The immunization study showed that the rRVGP produced in the bioreactor was of high immunogenic quality.pt_BR
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)pt_BR
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)pt_BR
dc.format.extentp. 4773-4783pt_BR
dc.language.isoEnglishpt_BR
dc.relation.ispartofApplied Microbiology and Biotechnologypt_BR
dc.titleDROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoproteinpt_BR
dc.typeArticlept_BR
dc.identifier.doi10.1007/s00253-018-8962-0pt_BR
dc.identifier.urlhttp://dx.doi.org/10.1007/s00253-018-8962-0pt_BR
dc.contributor.externalUniversidade Federal de São Carlos (UFSCar)¦¦Brasilpt_BR
dc.contributor.externalUniversidade Estadual Paulista Júlio de Mesquita Filho (UNESP)¦¦Brasilpt_BR
dc.identifier.citationvolume102pt_BR
dc.identifier.citationissue11pt_BR
dc.subject.keywordRabies virus glycoproteinpt_BR
dc.subject.keywordRabies vaccinept_BR
dc.subject.keywordDrosophila melanogaster S2pt_BR
dc.subject.keywordWAVE Bioreactorpt_BR
dc.subject.keywordScale-uppt_BR
dc.subject.keywordRecombinant protein productionpt_BR
dc.relation.ispartofabbreviatedAppl Microbiol Biotechnolpt_BR
dc.identifier.citationabntv. 102, n. 11, p. 4773-4783, jun. 2018pt_BR
dc.identifier.citationvancouver2018 Jun;102(11):4773-83pt_BR
dc.contributor.butantanAstray, Renato Mancini|:Pesquisador|:Lab. Biotecnologia Viral|:pt_BR
dc.contributor.butantanVentini, Daniella Cristina|:Técnico|:Lab. Biotecnologia Viral|:pt_BR
dc.contributor.butantanJorge, Soraia Attie Calil|:Pesquisador|:Lab. Biotecnologia Viral|:pt_BR
dc.contributor.butantanPereira, Carlos Augusto|:Pesquisador|:Lab. Biotecnologia Viral|:pt_BR
dc.sponsorship.butantanConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)¦¦402439/2013-9pt_BR
dc.sponsorship.butantanCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)¦¦064922/2014-01pt_BR
dc.identifier.bvsccBR78.1pt_BR
dc.identifier.bvsdbIBProdpt_BR
dc.description.dbindexedYespt_BR
item.openairetypeArticle-
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item.grantfulltextembargo_29990101-
item.languageiso639-1English-
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