Abstract
Messenger RNA (mRNA) based vaccines were instrumental in accelerating the end of the SARS-CoV-2 pandemic and are being aggressively developed as prophylaxes for a range of viral diseases. The swift adoption of mRNA-based therapeutics has also left open vast areas of opportunity for improving the development of mRNA-based drugs. One such area with immense potential focuses on the mRNA drug substance production, where mRNA is generated by a cell-free reaction called in vitro transcription (IVT). Process analytical technologies (PAT) are integral to the pharmaceutical industry and are necessary to facilitate agile process optimization and enhance process quality, control, and understanding. Due to the complexity and novelty inherent to the IVT reaction, there is a need for effective PAT that would provide in-depth, real-time insight into the reaction process to allow delivery of novel mRNA vaccines to patients faster in a more cost-effective way. Herein, we showcase the development of flow-nuclear magnetic resonance (flow-NMR) as a highly effective process-analytical tool for monitoring mRNA IVT reactions to support process development, optimization, and production.
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