posted on 2024-01-08, 14:36authored byMark S. Lowenthal, Abigail S. Antonishek, Karen W. Phinney
Lipid
nanoparticle-encapsulated mRNA (LNP-mRNA) holds great promise
as a novel modality for treating a broad range of diseases. The ability
to quantify mRNA accurately in therapeutic products helps to ensure
consistency and safety. Here, we consider a central aspect of accuracy,
measurement traceability, which establishes trueness in quantity.
In this study, LNP-mRNA is measured in situ using a novel liquid chromatography–mass
spectrometry (LC-MS) approach with traceable quantification. Previous
works established that oligonucleotide quantification is possible
through the accounting of an oligomer’s fundamental nucleobases,
with traceability established through common nucleobase calibrators.
This sample preparation does not require mRNA extraction, detergents,
or enzymes and can be achieved through direct acid hydrolysis of an
LNP-mRNA product prior to an isotope dilution strategy. This results
in an accurate quantitative analysis of mRNA, independent of time
or place. Acid hydrolysis LC-MS is demonstrated to be amenable to
measuring mRNA as both an active substance or a formulated mRNA drug
product.