RNA Leaders EU
Chromatography in mRNA Production: From Analytics to Purification
Speaker: Rok Sekirnik, Head of PC2, Process Development mRNA I pDNA
The talk will present case studies of leveraging the power of at-line HPLC monitoring for process development to increase purity and yield of mRNA production. Approaches to optimize plasmid purification and linearization, IVT reaction and mRNA purifi cation will be described, including purification methods to achieve >95% supercoiled plasmid purity, optimize linearization reaction, and increase of IVT reaction yield from 3-5 mg/mL to 10-12 mg/mL. Further, use of at-line anayltics to monitor automated, continuous production in single use bioreactors to produce multi-gram amounts of mRNA will be demonstrated. Oligo dT-based affinity purification will be described with approaches to optimize dynamic binding capacity to reach up to 6 g/L. Multimodal chromatography and reverse-phase chromatography cases will be discussed for capture of non-polyadenylated RNA, and removal of dsRNA.