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Synthetic, enzymatically produced DNA for gene therapy and vaccine applications

D Entiriwaa(1) Z Whiffen(1) A Walker(1) H Lanckriet(1) A Picher(1)


The manufacture of high-quality, GMP-grade DNA, crucial for gene therapy and vaccine applications, faces significant challenges including capacity limitations and complex sequence propagation issues. 4basebio has developed a proprietary, scalable, fully enzymatic process via our Trueprime™ amplification technology, yielding DNA at 1g/L; several orders of magnitude higher than plasmid fermentation yields. Our platform surpasses conventional plasmid fermentation, promising high-quality DNA with minimal footprint and turnaround time.  

4basebio’s  process is size and sequence independent, ensuring high yield and purity in less than a week. 4basebio easily produces complex sequences including ITRs and homopolymeric sequences including long polyA tracts, without risk of deletion or recombination. Unlike plasmid DNA, 4basebio DNA eliminates contamination from endotoxins or host proteins, and excludes bacterial sequences such as antibiotic resistance genes. 

Four types of application specific DNA are offered. opDNA™, optimized for mRNA production, enhances yields without the need for enzymatic linearization. Proinflammatory cytokine/chemokine levels in human PBMCs and in vivo protein expression are comparable to mRNA produced from linearised plasmid.  

hpDNA™ can be used in the production of AAV, replacing conventional plasmid triple-transfection. Using hpDNA™, viral genome titres, Full:Empty ratios and infectivity are equivalent to plasmid controls, across a range of serotypes. 

Finally, Hermes™, 4basebio's proprietary non-viral delivery system, encapsulates various payloads and can target specific cells or tissues. It offers long-term stability at 4°C, eliminating cold chain requirements. The combination of 4bb DNA or mRNA and Hermes™ nanoparticles can greatly accelerate the therapeutic development of gene therapy and vaccine programmes. 

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