Production of rabies VLPs in insect cells by two monocistronic baculoviruses approach

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dc.contributor(LABV) Laboratório de Biotecnologia Viralpt_BR
dc.contributorLab. Biologia Estruturalpt_BR
dc.contributor.authorBernardino, Thaissa Consonipt_BR
dc.contributor.authorAstray, Renato Mancinipt_BR
dc.contributor.authorPereira, Carlos Augustopt_BR
dc.contributor.authorBoldorini, Vera Luciapt_BR
dc.contributor.authorAntoniazzi, Marta Mariapt_BR
dc.contributor.authorJared, Simone Gonçalves Silvapt_BR
dc.contributor.authorNúñez, Eutimio Gustavo Fernándezpt_BR
dc.contributor.authorJorge, Soraia Attie Calilpt_BR
dc.identifier.citationBernardino TC, Astray RM, Pereira CA, Boldorini VL, Antoniazzi MM, Jared SGS, et al. Production of rabies VLPs in insect cells by two monocistronic baculoviruses approach. Mol. Biotechnol.. 2021 July;63:1068–1080. doi:10.1007/s12033-021-00366-z.pt_BR
dc.description.abstractRabies is an ancient zoonotic disease that still causes the death of over 59,000 people worldwide each year. The rabies lyssavirus encodes five proteins, including the envelope glycoprotein and the matrix protein. RVGP is the only protein exposed on the surface of viral particle, and it can induce immune response with neutralizing antibody formation. RVM has the ability to assist with production process of virus-like particles. VLPs were produced in recombinant baculovirus system. In this work, two recombinant baculoviruses carrying the RVGP and RVM genes were constructed. From the infection and coinfection assays, we standardized the best multiplicity of infection and the best harvest time. Cell supernatants were collected, concentrated, and purified by sucrose gradient. Each step was used for protein detection through immunoassays. Sucrose gradient analysis enabled to verify the separation of VLPs from rBV. Through the negative contrast technique, we visualized structures resembling rabies VLPs produced in insect cells and rBV in the different fractions of the sucrose gradient. Using ELISA to measure total RVGP, the recovery efficiency of VLPs at each stage of the purification process was verified. Thus, these results encourage further studies to confirm whether rabies VLPs are a promising candidate for a veterinary rabies vaccine.pt_BR
dc.description.sponsorship(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológicopt_BR
dc.description.sponsorship(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulopt_BR
dc.description.sponsorship(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorpt_BR
dc.relation.ispartofMolecular Biotechnologypt_BR
dc.rightsRestricted accesspt_BR
dc.titleProduction of rabies VLPs in insect cells by two monocistronic baculoviruses approachpt_BR
dc.contributor.external(USP) Universidade de São Paulopt_BR
dc.subject.keywordVirus-like Particlespt_BR
dc.subject.keywordInsect cellspt_BR
dc.relation.ispartofabbreviatedMol. Biotechnol.pt_BR
dc.identifier.citationabntv. 63, p. 1068–1080, jul. 2021pt_BR
dc.identifier.citationvancouver2021 July;63:1068–1080pt_BR
dc.contributor.butantanBernardino, Thaissa Consoni|:Desvinculado|:Lab. Biotecnologia Viral|:PrimeiroAutorpt_BR
dc.contributor.butantanAstray, Renato Mancini|:Pesquisador|:Lab. Biotecnologia Viralpt_BR
dc.contributor.butantanPereira, Carlos Augusto|:Pesquisador|:Lab. Biotecnologia Viralpt_BR
dc.contributor.butantanBoldorini, Vera Lucia|:Desvinculado|:Lab. Biotecnologia Viralpt_BR
dc.contributor.butantanAntoniazzi, Marta Maria|:Pesquisador:Docente PPGTox|:Lab. Biologia Estruturalpt_BR
dc.contributor.butantanJared, Simone Gonçalves Silva|:Desvinculado|:Lab. Biologia Estruturalpt_BR
dc.contributor.butantanJorge, Soraia Attie Calil|:Pesquisador|:Lab. Biotecnologia Viral|:Autor de correspondênciapt_BR
dc.sponsorship.butantan(CNPq) Conselho Nacional de Desenvolvimento Científico e Tecnológico¦¦152538/2012–7pt_BR
dc.sponsorship.butantan(FAPESP) Fundação de Amparo à Pesquisa do Estado de São Paulo¦¦2009/08038–1pt_BR
dc.sponsorship.butantan(CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior¦¦pt_BR
dc.sponsorship.butantanFundação Butantan¦¦pt_BR
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