The rapid advancement of mRNA therapeutics, driven by the success of mRNA-based vaccines, has created an urgent need for robust analytical methods to define and control the critical quality attributes (CQAs) of mRNA drug substances and drug products. Liquid chromatography (LC) methods—including ion-pair reverse-phase (IP-RP) HPLC, ion-exchange chromatography (IEX), and size-exclusion chromatography (SEC)—have emerged as powerful tools for the characterization of mRNA molecules. These techniques enable sensitive, high-resolution separation and quantification of mRNA-related impurities such as truncated transcripts, double-stranded RNA (dsRNA), and residual DNA or enzymes.

This presentation highlights a comprehensive LC-based analytical framework for evaluating the identity, integrity, purity, and stability of mRNA. Key case studies will demonstrate method development and performance in both early-phase development. Additionally, the integration of LC methods with orthogonal techniques such as UV, fluorescence, and mass spectrometry will be discussed to provide a more complete picture of mRNA quality. The role of these LC methods in supporting regulatory expectations and ensuring product consistency throughout the product lifecycle will also be reviewed.

Learning Objectives: 

1. Discuss the critical quality attributes (CQAs) of mRNA-based therapeutics and their implications for product quality and safety.

2. Explore the application of LC techniques (IP-RP, IEX, SEC) for characterization of mRNA integrity, purity, and impurities.

3. Gain insights into current regulatory trends and expectations for CQA monitoring of mRNA products.