Segmented Recombinant Vaccine Plasmids for Variant-Responsive Immunity
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Solution Overview
Problem
Current COVID-19 vaccines induce low levels of neutralizing antibodies (nAbs) that fail to control viral replication and prevent mild to moderate disease caused by Omicron variants, necessitating booster doses, and there is a need for a DNA-based platform that induces a strong protective immune response mediated by T cells to rapidly adapt vaccines against SARS-CoV-2 and its variants.
Innovation Solution
Development of a vaccine platform consisting of two plasmids, pCTV1 and pCTV2, derived from a commercial plasmid (pCDNA 3.1), optimized to express essential genes for mammalian cells, including CMV and SV40 promoters, multiple cloning sites, and bovine growth hormone polyadenylation signals, reducing size to 3.0-3.4 kb, which induce robust T cell responses and nAbs against SARS-CoV-2 and Influenza.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current COVID-19 vaccines are used, then vaccination coverage is achieved, but neutralizing antibody levels are low and fail to control viral replication of Omicron variants
Solution Approach 1:
The vaccine platform is segmented into two separate plasmids (pCTV1 and pCTV2), each carrying specific antigen genes. This segmentation allows for optimized expression of individual antigens and facilitates rapid adaptation to different variants by simply replacing the gene insert while maintaining the proven plasmid backbone structure that elicits strong T cell responses.
Solution Approach 2:
The plasmid structure has been modified with specific parameter changes including the incorporation of CMV and SV40 promoters, multiple cloning sites, and bovine growth hormone polyadenylation signals. These parameter changes optimize gene expression in mammalian cells and enhance the immunogenicity of the vaccine, producing high levels of neutralizing antibodies and robust T cell responses.
2Productivity
If plasmid size is reduced to 3.0-3.4 kb, then manufacturing efficiency and distribution are improved, but gene expression capability must be maintained
Solution Approach 1:
Non-essential sequences have been extracted and removed from the commercial plasmid pCDNA 3.1 to reduce the overall size to 3.0-3.4 kb. This streamlining improves manufacturing efficiency and distribution while the essential expression elements (CMV/SV40 promoters, MCS, polyA signals) are retained and optimized to maintain robust gene expression capability in mammalian cells.
Solution Approach 2:
The plasmid design applies local quality optimization by concentrating essential expression elements at specific locations: CMV and SV40 promoters at the 5' end, multiple cloning sites in the middle for flexible antigen insertion, and bovine growth hormone polyadenylation signals at the 3' end. This localized optimization ensures high expression efficiency within the reduced overall plasmid size.
Data Source
AI summary
Recombinant plasmids and immunogenic DNA compositions including a recombinant plasmid which carries the gene sequence encoding the Spike protein or the gene sequence encoding the NP protein, with the capacity to prevent high viral loads and moderate and severe clinical forms of the disease by stimulating the immune system, both against the ancestral strain of SARS-CoV-2 and influenza strains against different variants of concern that are currently circulating are disclosed. A bivalent recombinant plasmid and a immunogenic composition including the bivalent recombinant plasmid which carries the gene sequence encoding the NP and Spike protein, with the capacity to prevent high viral loads and moderate and severe clinical forms of the disease by stimulating the immune system, both against the influenza strain and SARS-COV-2 strain against different variants of concern that are currently circulating are disclosed.