Quality by design in mRNA manufacturing: at-line monitoring coupled with DOE for higher IVT yield and efficiency of purification

Tuesday 08:00 PST / 11:00 EST / 16:00 GMT / 17:00 CET

Sponsor

Quality by design in mRNA manufacturing: at-line monitoring coupled with DOE for higher IVT yield and efficiency of purification

Due to the catalytic, multicomponent nature of the IVT reaction, optimization is a multi-factorial problem, ideally suited to a design of experiments (DOE) approach for optimization and identification of design space. This webinar will describe how a data-driven model of process yield (g/L), including impact of nucleoside triphosphate (NTP) concentration and Mg:NTP ratio on reaction yield, can be derived to optimize reaction cost drivers (e.g., RNA polymerase and DNA template), while minimizing dsRNA formation. With optimal conditions, the yield of the IVT reaction can reach 25 g/L in batch.

This webinar will also describe how a high-throughput purification train optimization can be performed by coupling multiparallel (96 well) and spin-based purification devices at mg-scale, with DOE methodology. mRNA purification is achieved with affinity chromatography selective for polyadenylated mRNA (Oligo dT) coupled with reverse-phase chromatography to remove IVT components (NTPs, DNA, T7), and IVT byproducts, in particular dsRNA, a major immunogenic impurity. Elimination of dsRNA improves translation and minimizes the activation of innate immune response. With the advent of personalized, mRNA-based therapies, such as neoantigen mRNA vaccines, which require multiple mg doses, minimization of innate immune response may be critical to clinical success of mRNA therapeutics.

Use of at-line analytics to monitor IVT reaction
Coupling PAT with DOE to increase yield of IVT up to 25 g/L
Use of affinity chromatography for isolation of mRNA
Removal of dsRNA in scalable fashion

Rok Sekirnik

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

Rok Sekirnik graduated with a degree in chemistry from the University of Oxford, where he also completed his PhD. He has worked in the areas of mAb, vaccine and gene therapy vector development in various roles, including Principal Scientist at Novartis and Project Manager at Batavia Biosciences. His team at BIA Separations is focused on developing high-yielding, cost-effective purification approaches for pDNA and mRNA production.