Cumberbatch, Nathan
(2021)
The Physical and Chemical Origins of Amyloid at Interfaces.
Doctor of Philosophy thesis, University of Liverpool.
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Abstract
Amyloidosis is of serious concern within the modern world with an ageing global population, there is a significant drive to understand how amyloid forms in order to research and develop new therapeutics to combat this threat. This thesis proposes using a cross-disciplinary collaborative approach to determine multiple aspects within the amyloid-like fibrillation pathway for human insulin (HI) at acidic and neutral pH conditions. Chapter 2 uses Reflection Anisotropy Spectroscopy to obtain fibrillar orientation information with the goal to investigate whether structural morphology can direct fibril growth. Lack of reproducibility at the surfaces and the observation of a ‘blue-haze’ on the silicon wafers led to the method development in Chapter 3. Chapter 4 uses conventional biophysical techniques, fluorescence spectroscopy and electron microscopy to study hydrophobic functionalised mesoporous silica microparticles provides a scaffold for human insulin fibrillation to occur. These results show that the microparticles induced fibril morphology changes and inhibited or enhanced the fibrillation process, with respect to the human insulin control. Chapter 5 explores the mass and structural changes of adsorbed human insulin at a hydrophobic surface, by QCM-D and Raman spectroscopy. The results of which showed variable human insulin adsorption to the two differing hydrophobically functionalised surfaces at pH 2 and pH 7 conditions. Chapter 6 is the result of a collaboration with the Department of Physics and provides the first images obtained from infrared scanning near-field optical microscopy in reflection with a quantum cascade laser source of human insulin adsorbed to gold surfaces to obtain both structural and locational information. These preliminary results provide exciting opportunities for future development of this technique at other surfaces and proteins. Chapter 7 uses fluorescence excitation – emission matrices to uncover the origin of the deep-blue autofluorescence phenomenon, with human insulin, α-synuclein and other peptides.
| Item Type: | Thesis (Doctor of Philosophy) |
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| Divisions: | Faculty of Science and Engineering |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 26 Mar 2021 09:48 |
| Last Modified: | 18 Jan 2023 22:56 |
| DOI: | 10.17638/03117042 |
| Supervisors: |
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| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3117042 |
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