Optimization of mRNA vaccine candidate column chromatography using high throughput screening | Poster Board #111

In the process development space, there is an ongoing effort to identify effective approaches for optimizing mRNA vaccine candidate column chromatography. Many purification workstreams for mRNA-based therapeutics rely on hybridization-affinity chromatography, where the polyadenylated tail base pairs with an immobilized oligo-deoxythymidilic acid (OligodT) ligand. Although the industry has seen substantial advancements using hybridization chromatography, there remains further scope for establishing a platform process that can accommodate a variety of mRNA constructs. In this study, two types of 96-well OligodT plates were used, a monolithic plate and a resin-based plate, to screen 10 distinct mRNA constructs ranging in size from 1,700 to 4,000 nucleotides. With this scaled-down approach, binding isotherm studies were completed following the Langmuir Adsorption Model to determine the maximum binding capacity of each construct to the OligodT ligand. On the monolithic plate, it was shown that constructs of various sizes can exhibit similar binding capacities, whereas the resin-based plate seemed to be more dependent on size. Additional range-finding experiments were conducted on a smaller selection of constructs to determine optimal binding conditions. Adjusting the buffering agent, salt, and pH helped to pinpoint ideal buffer compositions and ranges, ultimately maximizing yield and purity. This study showcases the practicality of using 96-well chromatographic plates to conduct high-throughput screening to optimize mRNA purification in a fraction of the time compared to traditional resin screening.