| Author | Pinheiro, Cristiane Garboggini Melo de | |
| Author | Pedrosa, Mayara de Oliveira | |
| Author | Teixeira, Naiara Carvalho | |
| Author | Ano Bom, Ana Paula Dinis | |
| Author | van Oers, Monique M | |
| Author | Oliveira, Geraldo Gileno de Sá | |
| Access date | 2016-07-26T12:35:57Z | |
| Available date | 2016-07-26T12:35:57Z | |
| Document date | 2016 | |
| Citation | 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. | pt_BR |
| ISSN | 1756-0500 | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/15122 | |
| Sponsorship | PDTIS-FIOCRUZ. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). Instituto Nacional de Ciência e Tecnologia de Doenças Tropicais (INCT-DT). | pt_BR |
| Language | eng | pt_BR |
| Publisher | BioMed Central | pt_BR |
| Rights | open access | pt_BR |
| Title | Optimization of canine interleukin-12 production using a baculovirus insect cell expression system | pt_BR |
| Type | Article | pt_BR |
| Abstract | Interleukin-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 dogs | pt_BR |
| Affilliation | 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, Brasil | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Bio-Manguinhos. Laboratório de Macromoléculas. Rio de Janeiro, RJ, Brasil | pt_BR |
| Affilliation | Wageningen University. Laboratory of Virology. Wageningen, The Netherlands | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, Brasil / Instituto Nacional de Ciência e Tecnologia de Doenças Tropicais (INCT-DT). Salvador, BA, Brasil | pt_BR |
| Subject | Interleukin-12 | pt_BR |
| Subject | Dog | pt_BR |
| Subject | Protein expression optimization | pt_BR |
| Subject | Baculovirus | pt_BR |
| e-ISSN | 10.1186/s13104-016-1843-7 | |
