mRNA Fragmentation and Quality Assessment Using Ion Pair Reverse-Phase Analytics

Ana Ferjančič Budihna, Anže Martinčič Celjar, Sergeja Lebar, Andreja Gramc Livk and Aleš Štrancar

Cell & Gene Therapy Insights 2023; 9(9), 1231–1247 | DOI: 10.18609/cgti.2023.161

Abstract:
Rapid advancement of mRNA technology, as a response to the COVID-19 crisis, prompted an increased need for precise analytical methods to support the fast-paced mRNA process development. Accurate and robust analytics are required to support modifications in the mRNA production process, protocols, raw materials, in vitro transcription reaction, purification methods, scale-up, or final formulation processes, to ensure high quality and safety of the final product. This Innovator Insight demonstrates the application of an ion pair reverse phase chromatographic analytical method as a robust analytical tool to determine mRNA fragmentation while also separating in vitro transcription components from the main product. The method’s efficacy is assessed through a comprehensive stability study of a mRNA standard at different temperatures. The chromatographic analytical results are compared to the ones obtained by the capillary gel electrophoresis, a well-established method for the analysis of fragmented mRNA.