Chromatography in mRNA Production From Analytics to Purification

Event: mRNA Applications in Discovery & Development
Date: June 20, 2023
Presenter: Rok Sekirnik, Sartorius BIA Separations

This talk presents case studies of leveraging the power of at-line HPLC monitoring for process development to increase purity and yield of mRNA production. It describes approaches to optimize plasmid purification and linearization, IVT reaction and mRNA purification, including purification methods to achieve more than 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 analytics to monitor automated, continuous production in single use bioreactors to produce multi-gram amounts of mRNA are demonstrated. Oligo dT-based affinity purification is described with approaches to optimize dynamic binding capacity to reach up to 6 g/L. Multimodal chromatography and reverse-phase chromatography cases are discussed for capture of non-polyadenylated RNA, and removal of dsRNA.