Critical Evaluation of Key Methods for RNA Quantification.

The approval of safe and effective lipid nanoparticles-encapsulated mRNA (LNP-mRNA) vaccines during the SARS-CoV-2 pandemic is catalysing the development of the next generation of mRNA therapeutics. Accurate characterisation methods are crucial for assessing the quality and efficacy of these complex formulations. Several analytical techniques exist for the quantification of the mRNA drug substance, including UV spectroscopy, capillary electrophoresis, high-performance liquid chromatography (HPLC), and digital PCR (dPCR). Here, we compare these methods, highlight their limitations, and discuss critical considerations for experimental design to improve the agreement between different analytical methods.