Large RNA, including messenger RNA (mRNA), has recently become a new prospective class of drug products. The potential of mRNA technology for rapid vaccine development has recently been highlighted by the successful development of mRNA vaccines. Current analytical methods to characterize RNA therapeutics are limited and the development of analytical methods for the analysis of large RNA (>1000 nucleotides), including mRNA drug products, is challenging.

Large RNAs are not suitable for direct intact MS analysis. Therefore, digestion of the RNA with appropriate RNase enzymes prior to LC-MS/MS methods for oligonucleotide identifications is required. In this session we demonstrate the development of specific RNase mapping workflows optimized for the analysis of large RNA, including mRNA therapeutics. High-throughput approaches have been developed in conjunction with automated software analysis that match fragment MS/MS sequence data to the native full sequence, resulting in high sequence coverage of the mRNA therapeutic.