Data for: Oligonucleotide mapping via mass spectrometry to enable comprehensive primary structure characterization of an mRNA vaccine against SARS-CoV-2

Oligonucleotide mapping via liquid chromatography mass spectrometry mass
spectrometry (LC-MS/MS) was recently developed to support development of
Comirnaty®, the world’s first commercial mRNA vaccine which immunizes
against the SARS-CoV-2 virus. Analogous to peptide mapping of therapeutic
protein modalities, oligonucleotide mapping described here provides direct
primary structure characterization of mRNA, through enzymatic digestion,
accurate mass determinations, and optimized collisionally-induced
fragmentation. Sample preparation for oligonucleotide mapping is a rapid,
one-pot, one-enzyme digestion. The digest is analyzed via LC-MS/MS with an
extended gradient and resulting data analysis employs semi-automated
software. In a single method, oligonucleotide mapping readouts include a
highly reproducible and completely annotated UV chromatogram with
>98% sequence coverage and a microheterogeneity assessment of 5´
terminus capping and 3´ terminus poly(A) tail length. Oligonucleotide
mapping was pivotal to ensure the quality, safety, and efficacy of mRNA
vaccines by providing: confirmation of construct identity and primary
structure and assessment of product comparability following manufacturing
process changes. More broadly, this technique may be used to directly
interrogate the primary structure of RNA molecules in general.