Expression of a Novel Fusion Recombinant Protein VP8-VP1 in Escherichia coli as a Rota-HAV Vaccine Candidate

Introduction: Rotaviruses (RV) and hepatitis A virus (HAV) are pathogens responsible for more than 2 million hospitalizations, especially in developing countries, due to transmission through the fecal-oral route.Currently, there are several FDA-approved RV and HAV vaccines available which are based on killed or attenuated viruses.However, these vaccines often have side effects and low efficacy in eliciting specific immunity.Therefore, the design of a vaccine based on a recombinant protein, composed of RV and HAV antigens seems essential.Methods: We used bioinformatics tools to design and analyze the properties, predict the structure and evaluate the function, immunogenicity, antigenicity, and truncated sequences of HAV VP1 and RV VP8 as a dual vaccine platform.The predicted epitopes were expressed as a recombinant protein in Escherichia coli BL21 where half of the VP1 protein was fused with the Rota protein VP8 using pET24a expression vector,.Results: The expressed protein was confirmed by SDS-PAGE and Western blotting.Subsequently, high-scale expression, purification, refolding and determination of the protein concentration (~2.4 µg/µl) were obtained.Conclusion: Upon completion of the future immunogenicity evaluation through injection into mice, the present fusion protein can potentially serve as a candidate for a recombinant vaccine against both RV and HAV infections.