LC-MS Considerations for Oligonucleotide Assays

2nd Annual Oligos, Oligos, Oligos!

A special 3-part Live Webinar series focused on Oligonucleotide Analysis from Waters Corporation and Collaborators.

Seminar Series Summary

Oligonucleotide therapeutics are helping the world address previously untreatable diseases, and their clinical pipeline continues to see strong growth. In various forms, including small interfering RNA (siRNA), Antisense Oligonucleotides (ASOs), CRISPR single guide RNA (sgRNA), and others, these nucleic acid modalities often incorporate a range of modified nucleotides for enhanced stability, and are increasingly conjugated to tissue-specific ligands (GalNAc, lipids, peptides, mAb’s, etc.) to effect targeted cell delivery. To advance their development, these molecules and their impurities must be fully characterized, and their stability and pharmacokinetic properties must be accurately assessed.

In this webinar series, Waters’ scientists and collaborators from Novo Nordisk, Eli Lilly, and Synthon will provide state-of-the-art updates on the tools and techniques used to both confirm their identity and purity, and to quantify their pharmacokinetic properties in biological systems.  Join us as we delve into the science of oligos and explore the power of LC and LC-MS based approaches to analyze them.

EPISODE 3

Addressing Oligonucleotide Non-Specific Adsorption in IP-RPLC-UV-MS 

• Speaker: Jan Fučík / Synthon

Learn about the challenges of oligonucleotide non-specific adsorption during IP-RPLC-UV-MS analyses. Unwanted binding of these negatively charged molecules to surfaces in the chromatographic fluidic path can lead to poor recovery, distorted peak shapes, carry-over, and increased sodium adduct formation. Today’s presentation will focus on column testing using AMPcP and adenosine, evaluation of the inherent bioinertness of different instrument systems, and a comparison of various HFIP sources to assess their impact on sodium adduct levels.

Leveraging Multi-Reflecting TOF MS Technology for Oligonucleotides: Confirmation & Impurities, Mapping and More.

• Speaker: Alexandre Gomes / Waters Corporation

With the ever-increasing complexity of oligonucleotide therapeutics, there is a growing need for more powerful analytical tools  for characterization. The new Xevo MRT mass spectrometer from Waters features high mass resolution, accuracy and speed and has been demonstrated as an excellent platform for oligonucleotide LC-MS analysis. This presentation will highlight examples of intact mass confirmation and impurity work, MS-based sequencing and enzyme-based RNA mapping obtained with the Xevo MRT, showcasing the instrument's performance in combination with complete workflows offered by Waters from sample preparation to data processing and reporting.    

Presenter: Jan Fučík (Development Analyst for Oligonucleotides, Synthon)

Jan obtained his Master’s degree in Chemistry for Medical Applications and his Ph.D. in Analytical and Environmental Chemistry from Brno University of Technology. He has more than six years of experience with a broad range of mass spectrometry techniques (LC-ESI-3D-IT, LC-ESI-SQ, LC-ESI-TQ, LC-APCI-TQ, LC-ESI-qTOF, and MALDI-TOF), with a strong focus on LC-MS method development, sample preparation (SPE, QuEChERS), and statistical data evaluation. His expertise includes both targeted and untargeted analysis of small molecules (such as pharmaceutical residues and monomers for polymer production) and, more recently, oligonucleotides through his work at Synthon in the R&D Analytical/Oligonucleotide Department. At Synthon, he is responsible for LC-UV-MS method development for the analysis of oligonucleotide therapeutics, addressing the day-to-day analytical challenges these complex molecules present.

Presenter: Alexandre Gomes (Principal Mass Spectrometry Application Specialist, Waters Corporation)

Alex Gomes is a Principal MS Applications Specialist at Waters Corporation in the Americas Mass Spec Demo Lab.'s biopharmaceutical group. With over 16 years working with mass spectrometry bio applications, Alex is experienced with protein, peptide and oligonucleotide analysis, including LC-MS for qualitative and quantitative applications and specialized techniques such as HDX and ion mobility. Prior to joining Waters, he earned his Ph.D. in Chemistry from a leading university and published articles in peer-reviewed journals, showcasing his expertise in areas such as structural proteomics, ion mobility and HDX.