SARS-CoV-2 multi-epitope subunit vaccine proof-of-concept derived from the insilico study with protein expression in Escherichia coli BL21

Abstract

The protein subunit vaccine is the most considerably developed SARS-CoV-2 vaccine, according to the WHO vaccine tracker in 2023. The acceleration of vaccine development in 2 years of eradicating the COVID-19 pandemic is attainable due to the role of bioinformatics. This paper aims to evaluate strategies for developing multi-epitope SARS-CoV-2 recombinant vaccines in silico with high protein expression in Escherichia coli vector plasmid. The study was conducted by analysing SARS-CoV-2 epitopes using immunoinformatic tools provided by IEDB, codon optimisation, rare codon analysis, plasmid design, and ribosomal binding site (RBS) analysis were analysed using RNA structure 6.4, gene cloning by E. coli DH5α and protein expression by E. coli BL21. Each epitope peptide candidate was linked to a flexible linker sequence (GGGGS). GelAnalyzer 19.1 was utilised to determine the protein band of SDS-PAGE. The immunoinformatic study obtained a multi-epitope of the recombinant SARS-COV-2 vaccine with 7 epitopes for HLA-I allele candidates and 4 for HLA-II. It is demonstrated that the candidate vaccine protein was successfully cloned in E. coli DH5α and expressed in E. coli BL21. The result of this study will benefit the development of a SARS-CoV-2 protein recombinant vaccine that is safe, affordable, and efficient to induce T-cell response.