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Turpin, J.T. (2025) Part A - Formulation of Lipid Nanoparticles with RNA-based Payload – Key Considerations for Characterisation. Part B - Formulation and Biophysical Characterisation of Poly (A)-loaded Lipid Nanoparticles – Effects of Recombumin® on Physiochemical Properties. MRes thesis, University of Nottingham.

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Abstract

Part A - Lipid nanoparticles are latest lipid-based drug delivery systems that have recently drawn attention for their use in vaccine preparations by moderna and Pfizer-BioNTech for the transport of mRNA during COVID-19 pandemic. LNPs allow for the transport of delicate drug payloads, which degrade or incite immunogenic responses during circulation at normal physiological levels. This literature review aims to identify possible techniques for preparation and characterisation of LNPs carrying RNA based payload.

The most common types of LNPs for RNA based cargo are solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs). Both are suitable for transport. SLNs can carry both hydrophilic and hydrophobic drugs. NLC in comparison have greater biocompatibility due to a flexible membrane formed through structural tuning of surfactants used.

Surfactants such as ionisable lipids, PEGylated lipids, phospholipids and cholesterol have a key role in the structural characteristics and biocompatibility of the resultant LNPs. The combination of these components can be done in a variety of microfluidic mixers such as staggered Herringbone microfluidic mixer, an example of passive mixers. Chaotic flow in these passive mixers and increased surface area allows easy diffusion and mixing of aqueous and organic phase. Production of LNPs with passive mixers is preferred when handling RNA based cargo, to prevent the degradation of cargo through external interactions. Stability of LNPs can be measured by light scattering to identify PDI and zeta potential and aggregation behaviour. These parameters are important to predict the application in drug delivery.

This review focuses on the recent development in LNP based research and production. It aims to provide a comprehensive overview of different LNP formulations and their characterisation methods.

Part B - Lipid nanoparticles (LNPs) are small (less than a micron), lipid-based entities surrounding nucleotide-based agents sauch as mRNA as seen in mRNA vaccine developed by Pfizer and BioNTech during COVID-19. Use of ionisable lipids, phospholipids, cholesterol and PEGylated lipids forms a stable complex, capable of shielding delicate cargo (e.g a nucleotide).

During this project, we used a staggered herringbone mixing chip to create LNPs. Herringbone chip provides chaotic mixing and provides uniform diffusion which leads to uniform LNP particle synthesis. The LNPs thus produced were tested for their biophysical characteristics using dynamic light scattering (DLS) to determine their hydro dynamic radius, Zeta (ζ)-potential to measure their surface charge and Size Exclusion Chromatography coupled with Multi-Angled Light Scattering (SECMALS) to calculate the molar mass of the samples produced. Recombinant human albumin (rHA) Recombumin® Prime and Recombumin® Elite, products of Sartorius Albumedix Limited, were added to the LNP samples to test for LNP interaction and determine their effect on the suitability of the assay to detect the LNP. Increase in LNP size and reduction in surface charge, with the addition of albumin indicated the influence of albumin on the hydrodynamical properties of LNPs. SEC-MALS based investigations gave an indication of some interactions between LNP and albumin. On that account, it was inferred that DLS and SECMALs provided suitable baseline information about the biophysical properties of LNPs.

However, given the time frame of this work, it was not identified if the interaction between the two moieties were real and if they would affect overall functionality of the LNP.

Item Type:	Thesis (University of Nottingham only) (MRes)
Supervisors:	Jiwani, S.J. Harding, S.H.
Keywords:	Lipid Nanoparticles, drug delivery systems, Dynamic Light Scattering, SEC-MALS, Staggered Herringbone, Poly (A)
Subjects:	R Medicine > RS Pharmacy and materia medica
Faculties/Schools:	UK Campuses > Faculty of Science > School of Biosciences
Item ID:	82749
Depositing User:	Turpin, Joseph
Date Deposited:	12 Dec 2025 04:40
Last Modified:	12 Dec 2025 04:40
URI:	https://eprints.nottingham.ac.uk/id/eprint/82749

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