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• W3201442944 abstract "<ns3:p><ns3:bold>Background:</ns3:bold> The SARS-CoV-2 pandemic is a global threat to humans and the world’s economy. Effective and safe vaccines against this virus are essential to control and eradicate the pandemic. The currently applied vaccines carry SARS-CoV-2 spike-protein mRNA/cDNA. These vaccines go through several cellular processes in the recipients for producing antigens. On the contrary, the SARS-CoV-2 RBD (receptor binding domain)-protein is an antigen. It will directly stimulate antibody production against SARS-CoV-2. Hence, we propose to produce SARS-CoV-2 RBD-protein as a fast acting, effective and safe vaccine. </ns3:p><ns3:p> <ns3:bold>Methods:</ns3:bold> We propose to reconstruct a plasmid carrying three types of DNA sequences: RBD cDNA, FP (fusion peptide) DNA and sfGFP(superfolder green fluorescent protein), cDNA creating the RBD-FP-sfGFP DNA within an <ns3:italic>orf </ns3:italic>(open reading frame). <ns3:italic>Escherichia coli,</ns3:italic> C2566H, transformed with the reconstructed plasmid will express RBD-FP-sfGFP fusion protein producing green fluorescent <ns3:italic>cfu </ns3:italic>(colony forming unit). The RBD-protein will be separated from the sfGFP using an FP specific enterokinase, and eluted by HIC (<ns3:italic>hydrophobic interaction chromatography</ns3:italic>), detected with a BioVision Elisa kit, and quantified by spectrophotometry at UV280<ns3:sub>nm</ns3:sub>. </ns3:p><ns3:p> <ns3:bold>Results:</ns3:bold> The plasmid reconstruct will carry amp<ns3:sup>r </ns3:sup>(ampicillin-resistant) gene as a selective marker and a T7 promoter controlling the expression of RBD-FP-sfGFP fusion protein. The transformed <ns3:italic>Escherichia coli</ns3:italic> will efficiently express the RBD-FP-sfGFP fusion protein. The highly efficient sfGFP fused within the RBD-FP-sfGFP will produce green fluorescent <ns3:italic>cfu</ns3:italic>. The RBD-FP-sfGFP protein extract from the green <ns3:italic>cfu,</ns3:italic> digested by enterokinase and separated by the HIC will produce pure RBD protein. </ns3:p><ns3:p> <ns3:bold>Conclusion: </ns3:bold>A positive BioVision ELISA test detects <10 pg RBD protein/ml of the sample. A larger sample of the purified RBD protein can be used as a vaccine following a standard formulation and safety protocols. Once administered, the RBD protein will stimulate antibody production against the SARS-CoV-2 virus. The RBD protein has no potential to recombine with human genome.</ns3:p>" @default.
• W3201442944 created "2021-09-27" @default.
• W3201442944 creator A5006277366 @default.
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• W3201442944 date "2021-09-20" @default.
• W3201442944 modified "2023-09-27" @default.
• W3201442944 title "A study protocol to prepare an RBD protein for vaccine against COVID-19" @default.
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• W3201442944 doi "https://doi.org/10.12688/f1000research.54738.1" @default.
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