A study protocol to prepare an RBD protein for vaccine against COVID-19

Background: The SARS-CoV-2 pandemic is a global threat to humans and the world’s economy. Effective safe vaccines against this virus are essential control eradicate pandemic. currently applied carry spike-protein mRNA/cDNA. These go through several cellular processes in recipients for producing antigens. On contrary, RBD (receptor binding domain)-protein an antigen. It will directly stimulate antibody production SARS-CoV-2. Hence, we propose produce RBD-protein as fast acting, effective vaccine. Methods: We reconstruct plasmid carrying three types of DNA sequences: cDNA, FP (fusion peptide) sfGFP(superfolder green fluorescent protein), cDNA creating RBD-FP-sfGFP within orf (open reading frame). Escherichia coli, C2566H, transformed with reconstructed express fusion protein cfu (colony forming unit). be separated from sfGFP using specific enterokinase, eluted by HIC (hydrophobic interaction chromatography), detected BioVision Elisa kit, quantified spectrophotometry at UV280nm. Results: ampr (ampicillin-resistant) gene selective marker T7 promoter controlling expression protein. coli efficiently highly efficient fused cfu. extract cfu, digested enterokinase pure Conclusion: A positive ELISA test detects <10 pg protein/ml sample. A larger sample purified can used vaccine following standard formulation safety protocols. Once administered, virus. has no potential recombine human genome.