Thesis
Engineering lipid nanoparticles for nucleic acid delivery : effects of manufacturing, composition, and payload on structure and performance
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2026
- Thesis identifier
- T18034
- Person Identifier (Local)
- 202279357
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Lipid nanoparticles (LNPs) are the most clinically advanced delivery systems for nucleic acid therapeutics, with established use in messenger RNA (mRNA) vaccines and growing relevance for RNA- and DNA-based medicines. Despite their success, the relationships among manufacturing conditions, formulation composition, nucleic acid payload, and biological performance are not fully understood. Improved understanding of these relationships is needed to support the development of robust and reproducible LNP formulations. This thesis examines how manufacturing parameters, lipid composition, and nucleic acid payload influence the physicochemical properties and in vitro and in vivo performance of LNPs. Reproducible LNPs were first established using a NanoAssemblr-based microfluidic platform, enabling systematic optimisation of key processing variables. Using this foundation, LNP size was controlled by adjusting the aqueous-to-organic phase ratio, demonstrating maintained mRNA expression across a wider size range (60–120 nm) than typically employed, with differing trends observed in vitro and in vivo. The role of lipid composition was assessed by comparing clinically used and proprietary ionisable lipids with various sterols. While formulations displayed broadly similar physicochemical characteristics, differences in expression and biodistribution were observed in vitro and in vivo. Ionisable lipid structure was identified as a primary contributor to LNP performance, whereas sterol choice had more modest effects. The impact of nucleic acid payload identity was then investigated using LNPs encapsulating mRNA, self-amplifying RNA, plasmid DNA, and defined combinations thereof. Distinct in vivo expression profiles were observed, reflecting payload-specific biology, while combined-payload formulations exhibited additive behaviour, enabling modulation of expression kinetics without altering particle composition. Finally, the scalability of LNP production was briefly evaluated using an alternative microfluidic platform, demonstrating comparable in vivo expression across a range of manufacturing flow rates. Overall, this thesis provides insight into the factors that shape LNP performance and supports the rational design of LNP-based nucleic acid delivery systems.
- Advisor / supervisor
- Perrie, Yvonne
- Rattray, Zahra
- Resource Type
- DOI
Relations
Items
| Thumbnail | Title | Date Uploaded | Visibility | Actions |
|---|---|---|---|---|
|
|
PDF of Thesis T18034 | 2026-08-03 | Public | Download |