Automated direct mRNA sequence mapping using online partial RNase T1 digestion and 2D LC-MS

This webinar highlights a fully automated 2D LC-MS workflow for rapid, high-coverage mRNA characterization. By integrating online partial RNase T1 digestion directly into a 2D LC setup, the method enables controlled, reproducible digestion with seamless transfer into ion-pair reversed-phase separation.

The result:

• 93–99% sequence coverage in <60 minutes
• No post-digest sample handling
• Automated multi-attribute monitoring (sequence identity, 5’ cap, poly(A) tail)

The 2D LC configuration is central to achieving speed, resolution, and robustness in mRNA analysis.

Key Learnings:

• How 2D LC integrates online RNase digestion
• Why 2D LC improves separation and sequence coverage
• Benefits of automation for reproducibility and throughput
• Enabling multi-attribute mRNA analysis in a single workflow

Who should attend?

• mRNA analytics & LC–MS scientists (analytical development, characterization): especially anyone doing mRNA sequence mapping, oligoribonucleotide ID, or improving coverage/speed with LC–MS workflows.
• Biopharma QC / QC method owners working on mRNA identity testing and routine attribute measurements where automation, robustness, and reproducibility matter.
• Process development / CMC / MSAT teams supporting mRNA modalities who need faster turnaround data on critical quality attributes (CQAs) like 5’ cap, poly(A) tail length/heterogeneity, and sequence identity.
• Platform/automation & high-throughput labs looking to reduce hands-on time and eliminate post-digest sample handling while increasing throughput.
• Chromatography / LC system specialists interested in 2D LC configuration, online enzymatic digestion integration, and ion-pair reversed-phase separations for challenging RNA samples.
• R&D leaders / assay strategists evaluating next-gen, multi-attribute monitoring approaches that consolidate multiple measurements into one automated 2D LC–MS method.

Speaker: Jessica Dale (PhD Student in Oligonucleotide Analysis, University of Sheffield)