RNA Therapeutics Conference
Chromatography in mRNA Production: From Analytics to Purification
Speaker: Rok Sekirnik, Head of Process Development, mRNA I pDNA
The talk will describe 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 purification 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. Use of 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.