Optimizing mRNA Production: Achieving 25 g/L mRNA Yield With Low dsRNA by Integrating PAT

Watch the webinar to learn how to:

• Achieve scalable preparation of high-quality DNA template
• Understand the use of at-line analytics to monitor the IVT reaction
• Couple PAT with DOE to increase yield of IVT to 25 g/L
• Use affinity chromatography for isolation of mRNA
• Remove dsRNA, residual DNA template and RNA fragments in scalable fashion

Speaker:

Rok Sekirnik, Head of Process Development for mRNA and pDNA at Sartorius BIA Separations

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Abstract:

mRNA production requires DNA template as a critical raw material. In the webinar, discover an efficient purification process comprising scalable continuous in-line lysis, anion exchange chromatography (AEX) capture, and selective hydrophobic interaction chromatography (SHIC) for polishing supercoiled (sc)DNA. At-line monitoring of scDNA linearization ensures complete conversion in minimal time, optimizing reagent consumption.

DNA template is used in IVT production, which can be scaled from micrograms to multi-grams with rapid at-line PATfix monitoring of nucleoside triphosphate (NTP) consumption and mRNA production. A data-driven model of process yield can be derived to optimize reaction cost drivers, and minimize dsRNA formation – a critical quality attribute in mRNA products.

Our speaker will demonstrate how to achieve IVT yields of 25 g/L in batch mode and how to design an efficient fed-batch IVT reaction. He will also show how to optimize purification at µg-scale using multiparallel (96 well) and spin-based devices in Oligo dT and SDVB chemistries, and scale these methods to multi-mg levels for clinical manufacturing.