Improved Analytics for mRNA Vaccines: Multiplexed Impurities Analysis and Rapid 5’ Capping Efficiency

In this proposed work, InDevR will achieve two major goals. First, develop and aim to launch a multiplexed mRNA vaccine impurities assay with a same-day time to result. Specifically, aim to build on the demonstrated success in using antibodies as capture molecules in a fluorescence-based microarray format to develop an assay for detecting a variety of potential impurities relevant to mRNA vaccine manufacturing, designed for InDevR’s existing VaxArray platform. The VaxArray mRNA vaccine impurities assay will be capable of identifying and measuring quantity of a variety of potential impurities encountered in the production of DNA plasmids (such as E. coli host cell proteins, antibiotic resistance marker proteins, linearization enzymes, and nucleases) and in the production of mRNA constructs from those DNA plasmids (such as linearization enzymes, T7 polymerase, nucleases, capping enzymes, and double-stranded RNA). For the second goal, InDevR will explore improvements to their existing 5’CapQ assay for the measurement of intact (cap to tail) mRNA that provides a novel mRNA integrity measurement. The current kit has generated substantial interest and performs well for certain applications, enabling accurate and precise measurements of the total amount of intact (cap to tail) mRNA against a well-characterized, sequence-matched standard. However, the availability of such a standard is limited for many potential customers. Therefore, the goal of this improvement is to develop a ‘universal’ quantification standard to accompany the kit to increase the commercial utility of the assay.