(251c) Dynamic Capture and Elution of mRNA with Affinity Membranes for Vaccine Production

Although mRNA-based vaccines are known for their protection against SARS-CoV-2, they have applications well beyond this disease. With efficacies >95%, mRNA vaccines are highly translatable, noninfectious, and unlikely to integrate into a patient’s genome. Further increasing the applicability and accessibility of RNA therapies depends on overcoming downstream purification processing limitations. Industry currently relies on diffusion limited chromatography which is inefficient with respect to residence time, reduces final product integrity, requires large footprint and thus, impacts process economics and efficiency. This work describes a convective-based oligo-dT affinity membrane separation as an improved option to reduce the residence time of downstream operations while allowing for continuous processing and high retention of folded mRNA.

As part of a global study investigating affinity membrane fabrication, performance and modeling of mRNA capture, this presentation will focus primarily on experimental quantification and optimization of affinity membrane effectiveness in terms of dynamic capture and elution of mRNA. The development of a High-Performance Liquid Chromatography (HPLC) adaptable method will be described, resulting in membrane binding capacities of ~4.0 mg/mL of Firefly Luciferase (Fluc)-mRNA, with near 100% recovery of bound nucleic acids, and strong maintenance of product integrity in eluent samples. Furthermore, several optimization studies will be reported involving ligand surface concentration, ligand type and length, and feed conditions to maximize the recovery of bound nucleic acids while balancing experimental cost (Fig. 1).

Overall, results indicate that the modified RC membranes provide an improved alternative to current industrial separations. The membrane system is able to leverage a convective nature, hence speed of operation, to provide high mRNA capacity offering attractive production economics, while maintaining recovery and purity across operational conditions. Ultimately, further optimization of the affinity system and scale-up to other formats such as hollow fibers will pave the way to increasing the accessibility and application range of exciting, mRNA-based therapeutics.