Author | Barbosa, Mayra M. F. | pt_BR |
Author | Kanno, Alex I. | pt_BR |
Author | Pancakova, Violeta | pt_BR |
Author | Gonçalves, Viviane M. | pt_BR |
Author | Malley, Richard | pt_BR |
Author | Faria, Leonardo P. | pt_BR |
Author | Leite, Luciana C. C. | pt_BR |
Access date | 2021-07-09T14:40:33Z | |
Available date | 2021-07-09T14:40:33Z | |
Document date | 2021 | |
Citation | BARBOSA, Mayra M. F. et al. Optimization of expression and purifcation of Schistosoma mansoni antigens in fusion with Rhizavidin. Molecular Biotechnology, p. 1-9, 24 June 2021. | pt_BR |
ISSN | 1073-6085 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/48088 | |
Sponsorship | FAPESP Grant
2017/24632-6 and Fundação Butantan. MMFB received a CAPES fellowship.
VP was registered in the EMJMD LIVE (Erasmus+Mundus
Joint Master Degree Leading International Vaccinology Education), cofunded
by the EACEA (Education, Audiovisual and Culture Executive
Agency, award 2015–2023) of the European commission and received
a scholarship from the EACEA. | pt_BR |
Language | eng | pt_BR |
Publisher | Springer | pt_BR |
Rights | open access | pt_BR |
Subject in Portuguese | Proteínas recombinantes | pt_BR |
Subject in Portuguese | Proteínas | pt_BR |
Subject in Portuguese | Esquistossomose | pt_BR |
Subject in Portuguese | Biotina | pt_BR |
Subject in Portuguese | Schistosoma mansoni | pt_BR |
Title | Optimization of expression and purifcation of Schistosoma mansoni antigens in fusion with Rhizavidin | en |
Type | Article | pt_BR |
DOI | 10.1007/s12033-021-00355-2 | |
Abstract | Schistosomiasis causes signifcant morbidity and mortality. Vaccine eforts to date indicate the need to increase the immunogenicity of Schistosoma antigens. The multiple antigen-presenting system, whereby proteins are genetically fused to rhizavidin and afnity linked to biotinylated templates, enables the generation of robust immune responses. The objective of this work was to express and purify the S. mansoni antigens, SmTSP-2 and SmCD59.2, in fusion with rhizavidin. The fusion with rhizavidin greatly decreased the expression level of rSmTSP-2, but not rSmCD59.2, and both were expressed in the insoluble fraction, requiring optimization of culture conditions. Evaluation of diferent E. coli strains and media showed that BL21-DE3 cultured in Terrifc Broth provided the highest expression levels of both proteins. Investigation of a range of time and temperature of induction showed that E. coli strains expressing rRzv:SmTSP-2 and rRzv:SmCD59.2 showed the highest protein production at 23 °C for 15 h. Recombinant proteins were purifed by a single step of afnity chromatography allowing isolation of these proteins in high concentration and purity. The optimization process increased fnal soluble protein yield of rRzv:SmTSP-2 by fourfold and rRzv:SmCD59.2 by tenfold, providing~20 mg/L of each protein. Optimized fusion protein production will allow antigen use in biotin–rhizavidin afnity platforms. | en |
Affilliation | Instituto Butantan. Laboratório de Desenvolvimento de Vacinas. São Paulo, SP, Brasil / Universidade de São Paulo. Programa de Pós‑Graduação Interunidades em Biotecnologia. São Paulo, SP, Brasil. | pt_BR |
Affilliation | Instituto Butantan. Laboratório de Desenvolvimento de Vacinas. São Paulo, SP, Brasil. | pt_BR |
Affilliation | Instituto Butantan. Laboratório de Desenvolvimento de Vacinas. São Paulo, SP, Brasil / Université Claude Bernard Lyon. Villeurbanne, France. | pt_BR |
Affilliation | Instituto Butantan. Laboratório de Desenvolvimento de Vacinas. São Paulo, SP, Brasil. | pt_BR |
Affilliation | Boston Children's Hospital. Division of Infectious Diseases. Boston, USA. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Laboratório de Biomarcadores e Inflamação. Salvador, BA, Brasil. | pt_BR |
Affilliation | Instituto Butantan. Laboratório de Desenvolvimento de Vacinas. São Paulo, SP, Brasil. | pt_BR |
Subject | Protein expression | en |
Subject | Protein purifcation | en |
Subject | Fusion proteins | en |
Subject | Rhizavidin–biotin system | en |
Subject | Schistosoma proteins | pt_BR |
e-ISSN | 1559-0305 | pt_BR |