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Title: Exploiting reversible interactions : hydrogels and protein cross-linkers
Author: Belsom, Adam
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2011
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A series of low molecular weight thermoreversible cystine hydrogelators were synthesised via solid-phase chemistry. Novel hydrogels were found to gelate at concentrations of < 2 mM using microwave super-heating. Benzoyl cystine amide derivative hydrogel, which could form at a concentration of 0.5 mM, equivalent to 0.022% w/w of gelator with respect to water (an incredible 111,000 molecules of water gelated per single molecule of gelator), was applied to cell culture of cervical cancer (HeLa) cells, which were found to distribute within the gel. Hydrogels were produced on a microarray format using a novel strategy involving deposition of hydrogel solutions by inkjet printing. The incorporation of fluorescent dye (Rhodamine B) into hydrogels provided a novel means for studying hydrogel morphology. Reversible boronate chemistry was implemented for the capture and release of proteins and peptides onto a solid-support as part of a modified peptide enrichment strategy. The strategy was proven following synthesis of hydroxamic acid and catechol modified peptides and a study of their interaction with solid-supported phenylboronic acid. NHS active ester affinity tags and cross-linkers were synthesised and applied to a 3D proteomics cross-linking analysis pipe-line. The introduction of a PEG unit led to a cross-linker with increased hydrophilicity and improved observation of both inter and intra-protein cross-links by mass spectrometry.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available