ASMS 2025 Digested Oligonucleotide IP-RPLC-MS/MS Characterization of mRNA Sequence, 5’ Cap, and 3’ PolyA Tail
Map mRNA sequence, cap, and tail in a single LC-MS/MS assay
About this Poster
This poster presents a novel, automated workflow for comprehensive characterization of therapeutic mRNA using digested oligonucleotide LC-MS/MS analysis. The approach enables simultaneous sequence mapping, 5’ cap quantitation, and PolyA tail length assessment in a single assay. Using eGFP mRNA as a model, the study demonstrates high sequence coverage, accurate isomer differentiation, and streamlined reporting—all powered by the integrated Byos software platform and IPRP separation with the Phenomenex bioZen Oligo column.
Key Learnings:
- Learn how to perform sequence-level mRNA mapping with >99% coverage using enzymatic digestion and high-resolution MS/MS.
- See how 5’ capping efficiency and cap intermediates can be quantified directly from LC-MS/MS data.
- Discover how to deconvolute and measure PolyA tail distributions using automated MS1 analysis.
