Messenger RNA chromatographic purification: advances and challenges

Area of Science:

• Biotechnology
• Pharmaceutical Sciences
• Molecular Biology

Background:

• Messenger RNA (mRNA) technology offers significant potential for vaccines and therapeutics due to its adaptability, speed, efficacy, and safety.
• mRNA drug purity is a critical determinant of therapeutic efficacy and patient safety.
• Current chromatographic purification methods face challenges due to mRNA's size, composition, instability, and similarity to by-products.

Approach:

• Comprehensive analysis of physicochemical property differences between mRNA and proteins.
• Detailed review of state-of-the-art chromatographic purification technologies for mRNA.
• Classification of purification methods based on physicochemical properties (size, charge, hydrophobicity) and chemical structures (backbone, bases, cap, poly-A tail).

Key Points:

• Challenges in mRNA purification stem from its large size, simple composition, instability, and by-product similarity.
• Purification strategies focus on exploiting differences in size, charge, hydrophobicity, and chemical structures.
• Efforts to reduce double-stranded RNA (dsRNA) by-products during in vitro transcription (IVT) and post-IVT purification are highlighted.
• Emerging circular mRNA (circRNA) purification using chromatography is also discussed.

Conclusions:

• Advanced chromatographic techniques are essential for achieving high-purity mRNA required for safe and effective therapeutics.
• Addressing challenges like dsRNA by-products is key to optimizing mRNA purification processes.
• This review provides guidance for Quality by Design (QbD) in mRNA downstream process development.