Thesis
Investigating the critical quality attributes of mRNA-loaded lipid nanoparticles and their biomolecular corona using novel analytical pipelines
- Creator
- Rights statement
- Awarding institution
- University of Strathclyde
- Date of award
- 2026
- Thesis identifier
- T17970
- Person Identifier (Local)
- 202267049
- Qualification Level
- Qualification Name
- Department, School or Faculty
- Abstract
- Lipid nanoparticles (LNPs) have emerged as a promising platform for messenger RNA (mRNA) encapsulation, as demonstrated by their pivotal role during the coronavirus disease 2019 (COVID-19) pandemic. Despite the availability of several mRNA vaccines on the market and ongoing clinical trials, thorough characterization of formulation Critical Quality Attributes (CQAs), particularly in terms of stability and interactions with biological systems forming the biomolecular corona remains a complex challenge. This thesis established an advanced and efficient separation and characterization platform to evaluate three LNP prototypes distinguished by their cationic/ionizable lipid components, DOTAP, MC3, and SM-102. Initially, the stability of the DOTAP-LNP prototype was assessed alongside the development of a characterization method employing Asymmetric Flow Field-Flow Fractionation combined with multiple in-line optical detectors (AF4-MD) in Chapter 2. Subsequent in vitro cytotoxicity assessments of this prototype together with existing literature indicated cytotoxic effects associated with DOTAP-LNPs. Based on these findings, the study proceeded with alternative prototypes, MC3 and SM-102-LNPs. Biomolecular corona studies of two LNP prototypes with Bovine Serum Albumin (BSA) revealed that while both LNPs exhibited altered attributes following biomolecular corona formation, the changes were more pronounced for MC3-LNPs compared to SM-102-LNPs in Chapters 3-4. To further investigate whether SM-102-LNPs could inhibit significant affinity to other biomolecules, their interaction with High-Density Lipoprotein (HDL) was examined in Chapter 5. This analysis revealed that biomolecular corona formation of SM-102-LNPs is biomolecule-specific. Downstream analysis of the separated fractions led to the development of a gentle isolation pipeline for effectively separating LNPs from associated biomolecules. Overall, this thesis has established a foundational framework for the separation and characterization of CQAs for mRNA-LNPs. The findings demonstrate the potential of employing orthogonal, high-resolution separation techniques in the early prototyping of mRNA-LNP formulations and their nano-bio interactions. Furthermore, this work highlights the necessity of establishing standardised, robust, and versatile analytical platforms, such as the one developed herein, to enable the rapid characterisation of mRNA-LNP formulations. Such approaches can support regulatory decision-making by providing optimised protocols based on orthogonal analytical techniques, thereby accelerating the development and translation of these delivery systems.
- Advisor / supervisor
- Rattray, Zahra
- Treacher, Kevin
- Perrie, Yvonne
- Capomaccio, Robin
- Resource Type
- DOI
- Funder
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PDF of thesis T17970 | 2026-04-27 | Public | Download |