The Optimisation of the Expression of Recombinant Surface Immunogenic Protein of Group B Streptococcus in Escherichia coli by Response Surface Methodology Improves Humoral Immunity
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Group B Streptococcus (GBS) is the leading cause of neonatal meningitis and a common pathogen in livestock and aquaculture industries around the world. Conjugate polysaccharide and protein-based vaccines are under development. The surface immunogenic protein (SIP) is a conserved protein in all GBS serotypes and has been shown to be a good target for vaccine development. The expression of recombinant proteins in Escherichia coli cells has been shown to be useful in the development of vaccines, and the protein purification is a factor affecting their immunogenicity. The response surface methodology (RSM) and Box–Behnken design can optimise the performance in the expression of recombinant proteins. However, the biological effect in mice immunised with an immunogenic protein that is optimised by RSM and purified by low-affinity chromatography is unknown. In this study, we used RSM for the optimisation of the expression of the rSIP, and we evaluated the SIP-specific humoral response and the property to decrease the GBS colonisation in the vaginal tract in female mice. It was observed by NI–NTA chromatography that the RSM increases the yield in the expression of rSIP, generating a better purification process. This improvement in rSIP purification suggests a better induction of IgG anti-SIP immune response and a positive effect in the decreased GBS intravaginal colonisation. The RSM applied to optimise the expression of recombinant proteins with immunogenic capacity is an interesting alternative in the evaluation of vaccines in preclinical phase, which could improve their immune response.
KeywordsGroup B streptococcus Response surface methodology Protein expression
The authors would like to thank Miguel Muñoz, Selene Espinosa, Magaly Barrientos and América Abarca for their technical assistance and Luis Vidal for editing the English text.
JJ, JS and DD-D performed all experimental procedures, data collection, analysis, interpretation and the writing of this paper. JS, DD-D and DS developed the experimental rSIP purification procedures and the ELISA procedure. RV performed the statistical analysis and interpretation. AMK and AEV analysed and interpreted the results. AEV designed and directed the experiments and wrote and edited this paper. All authors read and approved the final manuscript.
This research was supported by the Instituto de Salud Pública de Chile and Millenium Institute on Immunology and Immunotherapy.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
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