Analytical methods for characterizing RNA by LC-MS/MS

This presentation will illustrate how chromatographic parameters and Orbitrap Tribrid MS fragmentation modes can be integrated to obtain reproducible and structurally informative readouts of native RNA modifications.

Register Free: https://www.chromatographyonline.com/lcgc_w/ribonucleic

Event Overview:

The Central Dogma of Molecular Biology states that the genetic information stored in our DNA, is transcribed into an intermediate molecule, RNA, which is then translated into protein. There are three main types of RNA associated with transcription/translation: messenger RNA, ribosomal RNA, and transfer RNA. All RNAs in the biological system carry post transcriptional modifications; chemical functional groups added to the base, the ribose sugar, or both. These modifications can be as simple as a methylation, to more hyper modifications that have multiple enzymatic steps and co-factors in their biosynthetic pathway. The gold standard in analytical technology for identification and characterization of these modifications remains steadfast in the realm of liquid chromatography hyphenated to high resolution accurate mass spectrometry. The analysis of native RNA and its chemical modifications remain technically demanding, particularly for oligonucleotides in the 8–40 nt range. RP-based separation strategy that avoids ion-pairing reagents are in demand. Such systems were evaluated with and without sulfolane to assess retention, signal stability, and fragmentation efficiency on high resolution Thermo Scientific^TM Orbitrap^TM Tribrid^TM mass spectrometers. Additional recent results on the substrate specificity of RNase 4 and colicin E5, which delineate the useful boundaries of enzymatic fragmentation strategies for MS-based RNA analysis will be shown.

Key Learning Objectives:

1. Discuss how to obtain data to get a comprehensive view of RNA samples.
2. Focus on robustness, resolution, and the practical implications for IP-free conditions.
3. Learn how mass spec fragmentation modes can be integrated to obtain reproducible and structurally informative readouts of native RNA modifications.

Who Should Attend:

• This webinar is designed for RNA and epitranscriptomics researchers, bioanalytical scientists, and mass spectrometry practitioners who are seeking robust, chemically definitive approaches to characterize RNA and its modifications. Attendees will benefit from practical LC-MS/MS strategies applicable to basic research, core facility workflows, and therapeutic RNA development.
• LC-MS/MS users and analytical scientists interested in exploring RNA and RNA modification analysis as an emerging application area, even if RNA is not currently their primary research focus.

Speakers:

Robert Ross
Senior Product Application Specialist
Thermo Fisher Scientific

Dr. Robert Ross obtained his Ph.D. from the University of Cincinnati where he studied post transcriptional modifications in RNA using a liquid chromatography tandem mass spectrometer. After graduation, he joined the University of Cincinnati College of Medicine in the Department of Cancer Biology where he led the effort in developing a core facility focusing on metabolomic, lipidomic, and nucleic acid mass spectrometry. He joined Thermo Fisher Scientific in 2021 as a Senior Product Application Specialist with a focus on Nucleic Acid Mass Spectrometry, "Nucleomics."

Stefanie Kaiser
Full Professor
Institute of Pharmaceutical Chemistry,
Department of Chemistry, Biochemistry and Pharmacy,
Goethe University Frankfurt, Germany

Prof. Dr. Stefanie Kaiser obtained her PhD in Pharmaceutical Chemistry from Johannes Gutenberg University Mainz, where she studied RNA modifications using mass spectrometry–based approaches. She conducted postdoctoral research at the Massachusetts Institute of Technology with Peter C. Dedon, focusing on LC-MS/MS methods to analyze DNA modification dynamics in vivo. Since 2020, she is Full Professor of Pharmaceutical Chemistry at Goethe University Frankfurt. Her research centers on the quantitative and mechanistic analysis of RNA modifications, with a particular emphasis on nucleoside and oligonucleotide mass spectrometry and the epitranscriptome in human health and disease.

Register Free: https://www.chromatographyonline.com/lcgc_w/ribonucleic