Biomanufacturing for RNA Biologics

Learning Outcomes

After successfully completing this course, postgraduate students will be able to:

In terms of knowledge and understanding

• describe the structural and biochemical properties of RNA biologics relevant to their design, stability, and therapeutic function
• explain the upstream biomanufacturing operations for RNA biologics, including DNA template design and production and in vitro transcription processes at small and large scale
• describe downstream RNA purification strategies including tangential flow filtration and RNA-specific chromatography, and their application to mRNA therapeutic manufacturing
• identify established and emerging therapeutic applications of RNA biologics in gene therapy, cell therapy, and mRNA-based treatments

In terms of skills

• apply quantitative bioprocessing principles to analyse upstream and downstream unit operations for RNA biologic manufacturing
• evaluate and compare RNA purification strategies for yield, purity, and scalability in the context of mRNA therapeutic production
• critically analyse primary research and industry literature on RNA biomanufacturing and mRNA therapeutic development
• communicate biomanufacturing process designs and analyses through written reports and oral presentations
• integrate guest lecture content from academic and industry experts into a coherent understanding of current RNA biologic manufacturing practice
• apply knowledge of RNA biology and bioprocessing to evaluate current challenges and opportunities in the RNA therapeutics field
• work autonomously to synthesise information from multiple sources and develop evidence-based conclusions about RNA biomanufacturing

In terms of competencies

• integrate knowledge from RNA biology, bioprocess engineering, and pharmaceutical manufacturing to address challenges in RNA biologics production
• critically evaluate scientific and industrial literature and identify key advances and gaps in RNA biomanufacturing
• make informed decisions when assessing manufacturing strategies for novel RNA therapeutic modalities
• advance scientific thinking by engaging with cutting-edge research and industry perspectives through guest lectures and literature
• communicate scientific and technical findings on RNA biologics effectively to specialist and non-specialist audiences

Module Syllabus

1. Introduction to RNA biologics: types of therapeutic RNA (mRNA, siRNA, saRNA, circRNA), mechanisms of action, and clinical landscape
2. RNA structure, stability, and chemical modifications: secondary structure, modified nucleosides, and their role in immunogenicity and stability
3. DNA template design and production: plasmid design, linearisation, large-scale DNA template manufacturing for in vitro transcription
4. In vitro transcription (IVT): reaction principles, enzyme systems, scale-up from small-scale to large-scale IVT for mRNA production
5. RNA capping, polyadenylation, and quality control: co-transcriptional and enzymatic capping, poly-A tail addition, and analytical methods
6. Downstream processing I: RNA purification principles, precipitation, and filtration-based approaches including tangential flow filtration
7. Downstream processing II: chromatographic purification of RNA — oligo-dT, size exclusion, and ion exchange methods for mRNA therapeutics
8. RNA formulation and delivery: lipid nanoparticles, encapsulation efficiency, and considerations for storage and administration
9. Scale-up and GMP manufacturing of RNA biologics: regulatory considerations, process analytical technology, and quality systems
10. Applications I — mRNA therapeutics: vaccines (COVID-19 and beyond), protein replacement therapies, and cancer immunotherapy
11. Applications II — RNA-based gene and cell therapy: siRNA, antisense oligonucleotides, and RNA-guided genome editing strategies
12. Industry perspectives: guest lectures from academic and industry experts on synthesis, purification, formulation, and scale-up of RNA biologics

Suggested Bibliography

• RNA Therapeutics: The Evolving Landscape of RNA Therapeutics 1st Edition by Paloma H. Giangrande (Editor), Vittorio de Franciscis (Editor), John J. Rossi (Editor). Academic Press, 2022, ISBN-13: ‎978-0128215951
• RNA Vaccines Methods and Protocols, Springer Protocols, by Thomas Kramps (Editor), Knut Elbers (Editor), Humana Press, 2024
• Pardi, N. et al. mRNA vaccines — a new era in vaccinology. Nature Reviews Drug Discovery 17, 2018.
• Sahin, U. et al. mRNA-based therapeutics — developing a new class of drugs. Nature Reviews Drug Discovery 13, 2014.
• Course Lecture Notes, Slides (distributed via myCourses, McGill University’s learning management system)

Related academic journals:

Molecular Therapy, Nature Biotechnology, Journal of Controlled Release, Nucleic Acids Research, Biomaterials