Poster Presentation Australasian RNA Biology and Biotechnology Association 2024 Conference

mRNA Design and Production: A Streamlined Approach to Accelerate Vaccine and Therapy Development (#116)

Natasha Chaudhary , Hannah Tompkins , Abhishek Kulkarni , Helen Gunter , Magdalena Budzinska , Seth Cheetham , Tim Mercer

mRNA vaccines and therapies provide a new approach to treating diseases. However, traditional mRNA vaccine manufacturing from plasmid DNA is laborious, error-prone and harbours impurities. To overcome these challenges, we have developed a streamlined mRNA manufacturing method, that offers a high degree of flexibility and scalability to meet changing production needs.

mRNA manufacturability and stability depend on its design. Current mRNA design software uses codon optimisation, favouring the most abundant human codons1,2, but that doesn’t always ensure manufacturability. Our new software simplifies the mRNA design process, reducing codon-bias and ensuring mRNA manufacturability with minimal impact on putative translation rates and stability.

Instead of using plasmid DNA, our process employs a cell-free synthetic “gBlock” DNA template, which is amplified using PCR to produce in-vitro transcription templates. This approach significantly decreases template production time, effectively eliminates endotoxin contamination and prevents poly(A) tail truncation. Thus, this adaptable platform can rapidly manufacture various drug candidates, accelerating discovery and development cycles.

We have developed a pipeline that analyses the key quality attributes of mRNA and their biological activity and safety. We use long-read Oxford Nanopore Technologies (ONT) sequencing to comprehensively measure sequence identity, length, integrity, and purity for both mRNA and DNA templates3. We employ a cell-based reactogenicity assays to assess dsRNA or other impurities in our mRNA products. Additionally, we measure quality, stability and efficacy of our LNP-encapsulated mRNA through in-vitro and in-vivo expression analysis. Together, our pipeline shows that mRNAs manufactured using g-blocks are of higher quality and biological activity than their plasmid-templated counterparts.

Through manufacturing hundreds of mRNA vaccines and therapies, we continually refine our pipeline, driving innovation and solidifying our leadership in mRNA production and research.

 

 

  1. 1. Vostrosablin, N. et al. NAR Genom. Bioinf. 6, lqae028 (2024). 2. Zhang, H. et al. Nature 621, 396-403 (2023). 3. Gunter, H. M. et al. Nat. Commun. 14, 5663 (2023)