Using nanopore sequencing for mRNA vaccine quality control: a journey from R&D to GMP

Abstract The production of mRNA vaccines involves costly and time-intensive quality testing. Contaminants, including double-stranded, degraded, or off-target mRNAs, can undermine the efficacy and safety of the vaccine and may emerge at any stage of the mRNA manufacturing process. We describe the development and validation of an Oxford Nanopore sequencing pipeline, designed for quality control of mRNA vaccines throughout their manufacture. Our pipeline streamlines the assessment of mRNA sequence identity, purity, and integrity from the 5’ cap to the poly(A) tail. To adapt this analytical pipeline for clinical-grade mRNAs, we also describe the development of mRNA standards, suitable for the qualification and validation of quality control equipment and analytical methods. Our results demonstrate that Oxford Nanopore sequencing delivers comprehensive mRNA quality control information, thereby supporting routine mRNA vaccine manufacturing and facilitating continuous process improvements.  Biography Helen Gunter is a Senior Research Scientist at the BASE Facility, University of Queensland. Her research focuses on advancing tools to enhance the design, quality, and efficacy of mRNA vaccines.