Optimization of canine interleukin-12 production using a baculovirus insect cell expression system

Detalhes bibliográficos
Autor(a) principal: Pinheiro, Cristiane Garboggini Melo de
Data de Publicação: 2016
Outros Autores: Pedrosa, Mayara de Oliveira, Teixeira, Naiara Carvalho, Ano Bom, Ana Paula Dinis, van Oers, Monique M, Oliveira, Geraldo Gileno de Sá
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/15122
Resumo: PDTIS-FIOCRUZ. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Instituto Nacional de Ciência e Tecnologia de Doenças Tropicais (INCT-DT).
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spelling Pinheiro, Cristiane Garboggini Melo dePedrosa, Mayara de OliveiraTeixeira, Naiara CarvalhoAno Bom, Ana Paula Dinisvan Oers, Monique MOliveira, Geraldo Gileno de Sá2016-07-26T12:35:57Z2016-07-26T12:35:57Z2016PINHEIRO, C. G. M. et al. Optimization of canine interleukin-12 production using a baculovirus insect cell expression system. BMC Res Notes, v. 9, p. 36, 2016.1756-0500https://www.arca.fiocruz.br/handle/icict/1512210.1186/s13104-016-1843-7PDTIS-FIOCRUZ. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Instituto Nacional de Ciência e Tecnologia de Doenças Tropicais (INCT-DT).Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / CAPES. Programa de Pós‑graduação em Biotecnologia em Saúde e Medicina Investigativa. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilFundação Oswaldo Cruz. Bio-Manguinhos. Laboratório de Macromoléculas. Rio de Janeiro, RJ, BrasilWageningen University. Laboratory of Virology. Wageningen, The NetherlandsFundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia de Doenças Tropicais (INCT-DT). Salvador, BA, BrasilInterleukin-12 is an important cytokine in mediating cellular immune responses. Results: Recombinant single-chain canine IL-12 was produced in a baculovirus-insect cell system with the aim of conducting further studies on modulation of immune responses in dogs. To optimize the production of recombinant canine IL-12, a classical baculovirus and a modified vector (chitinase A and v-cathepsin knockout) were used containing a native or an optimized insert of canine IL-12. The optimized IL-12 construct contained the GP64 signal peptide and was synthesized with optimized codons for expression in Trichoplusia ni cells. Dot-blot and Western blot analysis showed the highest production levels of recombinant IL-12 protein by the use of the modified baculovirus vector containing the optimized insert, at a multiplicity of infection of five and at 48 h after infection. The recombinant cytokine was successfully purified and showed a good degree of purity, integrity, folding, and yield, with very little endotoxin contamination. Recombinant canine IL-12 induced IFN-γ in canine lymphocytes, indicating that it was biologically active. Conclusion: Therefore, this study describes an efficient method to produce adequate amounts of biologically active canine IL-12, useful for immunomodulation studies in dogsengBioMed CentralOptimization of canine interleukin-12 production using a baculovirus insect cell expression systeminfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleInterleukin-12DogProtein expression optimizationBaculovirusinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/15122/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALPinheiro CGM Optimization of canine....pdfPinheiro CGM Optimization of canine....pdfapplication/pdf1943114https://www.arca.fiocruz.br/bitstream/icict/15122/2/Pinheiro%20CGM%20Optimization%20of%20canine....pdf8e27eea6e438cb36e13f2ac80411d376MD52TEXTPinheiro CGM Optimization of 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dc.title.pt_BR.fl_str_mv Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
title Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
spellingShingle Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
Pinheiro, Cristiane Garboggini Melo de
Interleukin-12
Dog
Protein expression optimization
Baculovirus
title_short Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
title_full Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
title_fullStr Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
title_full_unstemmed Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
title_sort Optimization of canine interleukin-12 production using a baculovirus insect cell expression system
author Pinheiro, Cristiane Garboggini Melo de
author_facet Pinheiro, Cristiane Garboggini Melo de
Pedrosa, Mayara de Oliveira
Teixeira, Naiara Carvalho
Ano Bom, Ana Paula Dinis
van Oers, Monique M
Oliveira, Geraldo Gileno de Sá
author_role author
author2 Pedrosa, Mayara de Oliveira
Teixeira, Naiara Carvalho
Ano Bom, Ana Paula Dinis
van Oers, Monique M
Oliveira, Geraldo Gileno de Sá
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Pinheiro, Cristiane Garboggini Melo de
Pedrosa, Mayara de Oliveira
Teixeira, Naiara Carvalho
Ano Bom, Ana Paula Dinis
van Oers, Monique M
Oliveira, Geraldo Gileno de Sá
dc.subject.en.pt_BR.fl_str_mv Interleukin-12
Dog
Protein expression optimization
Baculovirus
topic Interleukin-12
Dog
Protein expression optimization
Baculovirus
description PDTIS-FIOCRUZ. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Instituto Nacional de Ciência e Tecnologia de Doenças Tropicais (INCT-DT).
publishDate 2016
dc.date.accessioned.fl_str_mv 2016-07-26T12:35:57Z
dc.date.available.fl_str_mv 2016-07-26T12:35:57Z
dc.date.issued.fl_str_mv 2016
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv PINHEIRO, C. G. M. et al. Optimization of canine interleukin-12 production using a baculovirus insect cell expression system. BMC Res Notes, v. 9, p. 36, 2016.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/15122
dc.identifier.issn.pt_BR.fl_str_mv 1756-0500
dc.identifier.eissn.none.fl_str_mv 10.1186/s13104-016-1843-7
identifier_str_mv PINHEIRO, C. G. M. et al. Optimization of canine interleukin-12 production using a baculovirus insect cell expression system. BMC Res Notes, v. 9, p. 36, 2016.
1756-0500
10.1186/s13104-016-1843-7
url https://www.arca.fiocruz.br/handle/icict/15122
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
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https://www.arca.fiocruz.br/bitstream/icict/15122/3/Pinheiro%20CGM%20Optimization%20of%20canine....pdf.txt
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