{"id":{"repo_id":"cambridge","oai_identifier":"oai:www.repository.cam.ac.uk:1810/318229"},"canonical_url":"https://search.dev.ndltd.org/etd/cambridge/oai:www.repository.cam.ac.uk:1810/318229","repository":{"repo_id":"cambridge","name":"Cambridge University","base_url":"https://api.repository.cam.ac.uk/server/oai/request"},"display":{"title":"Biophysical approaches to characterising protein-protein interactions and intermediate species in protein aggregation","abstract":"The phenomenon of protein misfolding and aggregation has been associated with over 50 human diseases, including Parkinson’s disease (PD). While the hallmark deposits in aggregation-associated diseases are primarily composed of fibrillar species, intermediate oligomeric species that form during the aggregation process are believed to be a major cause of toxicity. Such species are relatively poorly characterised due to several challenges that render them inaccessible to most conventional techniques; oligomers are only present at extremely low concentrations in the aggregation reaction, and are additionally highly heterogeneous and often transient in nature. In this thesis, I present complementary approaches to address these difficulties. Firstly, an ensemble of stable, kinetically trapped oligomers with varying biophysical characteristics was established, enabling detailed structure-toxicity relationships to be investigated. Secondly, a method for the simultaneous single-molecule level characterisation and fractionation of oligomeric species under native conditions was established and applied to studying intermediate species in Parkinson’s disease-associated protein aggregation. 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Firstly, an ensemble of stable, kinetically trapped oligomers with varying biophysical characteristics was established, enabling detailed structure-toxicity relationships to be investigated. Secondly, a method for the simultaneous single-molecule level characterisation and fractionation of oligomeric species under native conditions was established and applied to studying intermediate species in Parkinson’s disease-associated protein aggregation. Finally, I present a general approach to optimising experimental design, which maximises both the efficiency and information gain of experiments, and demonstrate its validation and application to several experimental systems.","abstract_has_math":false,"creators":["Xu, Catherine"],"institution":"University of Cambridge","degree_name":"Doctor of Philosophy (PhD)","degree_level":"Doctoral","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Knowles, Tuomas","Dobson, Christopher"],"committee_chairs":[],"committee_members":[],"year":2020,"date_issued":"2020-09-29","date_published":"2020-09-29","updated_at":"2026-07-22T22:24:17Z","subjects":["Protein aggregation","Amyloid","Oligomers","Alpha-synuclein","Microfluidics"],"languages":["eng"],"rights":[],"rights_urls":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/14d3dc54-efd7-4541-be62-5d4d0dd8a420/download","https://www.rioxx.net/licenses/all-rights-reserved/"],"identifier_entries":[]},"links":{"outbound_url":"https://doi.org/10.17863/CAM.65349","outbound_label":"DOI","outbound_source":"dc:identifier.doi"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Knowles, Tuomas","Dobson, Christopher"]},{"key":"dc:contributor.sponsor","label":"Sponsor","values":["Herchel Smith Fund"]},{"key":"dc:creator","label":"Author","values":["Xu, Catherine"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2020-09-29"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of Cambridge"]},{"key":"dc:relation.isreferencedby.uri","label":"Dc Relation Isreferencedby URI","values":["https://www.repository.cam.ac.uk/handle/1810/318229"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["Doctoral"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Protein aggregation","Amyloid","Oligomers","Alpha-synuclein","Microfluidics"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/14d3dc54-efd7-4541-be62-5d4d0dd8a420/download","https://www.rioxx.net/licenses/all-rights-reserved/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.doi","label":"DOI","values":["10.17863/CAM.65349"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/f2829d23-f685-4475-a4cf-ae167efd5f37/download"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["The phenomenon of protein misfolding and aggregation has been associated with over 50 human diseases, including Parkinson’s disease (PD). 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Secondly, a method for the simultaneous single-molecule level characterisation and fractionation of oligomeric species under native conditions was established and applied to studying intermediate species in Parkinson’s disease-associated protein aggregation. 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