LC-MS for RNA Therapeutics to Ensure Product Quality and Process Consistency

The development momentum for RNA modalities has accelerated with the recent approvals of mRNA-based vaccines and RNA(CRISPR)-based gene therapies. These approvals have highlighted the need for the development of robust analytical methods for monitoring the relevant critical quality attributes (CQAs) of RNA molecules to ensure proper control of manufacturing processes and product quality. Oligonucleotide mapping via liquid chromatography coupled to mass spectrometry (LC-MS) has now been established as an effective tool for RNA CQA assessment, however, the limited tool sets for efficient acquisition and processing of the LC-MS data from an RNA digest have presented a time-consuming challenge to the innovators bringing these early molecules to market.

In this webinar, we present a workflow for RNA CQA analysis developed on the waters_connect informatics platform. Data acquired on this compliant-ready platform was analyzed using the INTACT Mass application as the engine for data processing and producing RNA fragment assignments, assisted by two external microApps: mRNA Cleaver (to generate in silico fragments for targeted search), and Coverage Viewer (to visualize digestion data coverage). We will demonstrate these tools with application to CQA analysis of both sgRNA (ID and Structural Fidelity) and mRNA (ID, Structural Fidelity, 5’Capping, 3’ PolyA Tail A) molecules.

What you will learn from this presentation:

• A workflow for acquisition and data analysis from RNA digestion fragment mapping studies.
• Application of this workflow to sgRNA and mRNA CQAs.
• Discussion of the challenges and opportunities deploying these methods for routine analysis, including the use of the BioAccord LC-MS System and waters_connect informatics to make MS based RNA analysis accessible to non-MS experts.

Who should attend?

• Scientists, Technicians and Managers, who are charged with analyzing therapeutic RNA samples.
• Others from pharmaceutical and contract organizations looking towards developing and commercializing these molecules, who want to better understand the analytical challenges of producing high quality products and processes.

Presenter: Ying Qing Yu (Biopharmaceutical Program Science Team Leader, Waters Corporation)

Ying Qing is the Biopharmaceutical Program Science Team Leader at Waters Corporation. She joined Waters Corporation in 2001, shortly after she received her Ph.D. from the Analytical Chemistry Department at Purdue University. Her area of expertise is on analytical method development for various Biotherapeutics using UPLC/QTOF MS platform, including sample preparations, LC-MS optimization, and development of application specific software tools. Recently, her team has been taking on the challenges of developing LC-MS solutions for nucleic acid characterization.