Characterization and sequence mapping of large RNA and mRNA therapeutics using HRAM

The presentation showcases the innovative partial T1 digest strategy developed at the University of Sheffield for sequencing mRNA therapeutics. This approach, which utilizes LCMS, enables rapid and efficient sequence validation with over 90% coverage in less than 90 minutes. 

The technique involves generating overlapping fragments that are visualized through advanced mapping tools, providing clear, color-coded linear maps and spiral plots that indicate sequence coverage and confidence levels. This method has proven particularly valuable for large mRNA molecules, such as self-amplifying RNAs, which can be up to 678,000 nucleotides in length. The collaboration with manufacturing groups exemplifies the practical application of this technology in real-world scenarios, ensuring the integrity and efficacy of mRNA therapeutics. The presentation concludes by highlighting the establishment of a dedicated RNA facility at the University of Sheffield, which aims to further streamline and enhance mRNA analysis and characterization.

Learning points

• Critical Quality Attributes of mRNA: Understanding and monitoring the critical quality attributes (CQAs) of mRNA therapeutics and vaccines are essential.
• Advanced Sequencing Techniques: The use of advanced sequencing methods, such as Sanger sequencing and next-generation sequencing, is crucial for ensuring the correct identity of mRNA sequences.
• Impurity Monitoring and Safety: Monitoring impurities, such as double-stranded RNA and residual DNA from the manufacturing process, is vital for the safety of mRNA therapeutics.

Who should attend?

• Researchers and Scientists: Those working in molecular biology, biochemistry, and pharmaceutical sciences who are interested in the latest techniques for mRNA analysis and characterization.
• Biopharmaceutical Professionals: Professionals involved in the development and production of mRNA-based therapeutics and vaccines, particularly those focusing on quality control and regulatory compliance.
• Analytical Development Teams: Teams dedicated to improving analytical methods for validating the sequence and integrity of mRNA therapeutics.

Presenter:  Prof. Mark Dickman (Professor, University of Sheffield)

Prof. Dickman obtained a first-class honours degree in biochemistry/chemistry and his Ph.D. at the Krebs Institute, University of Sheffield. He then joined a biotechnology company, Transgenomic Ltd, and since 2003 has worked in the Dept. of Chemical and Biological Engineering at the University of Sheffield, where his research focuses on the development of analytical methods to characterize biomolecules including mRNA and oligonucleotide therapeutics.