{"id":{"repo_id":"cambridge","oai_identifier":"oai:www.repository.cam.ac.uk:1810/344696"},"canonical_url":"https://search.dev.ndltd.org/etd/cambridge/oai:www.repository.cam.ac.uk:1810/344696","repository":{"repo_id":"cambridge","name":"Cambridge University","base_url":"https://api.repository.cam.ac.uk/server/oai/request"},"display":{"title":"Engineering of an anti-PAI-1 antibody with tissue specific activation","abstract":"Antibody-based therapies can have shortcomings if on-target activity causes dose-limiting toxicities. A pro-antibody, inspired by naturally occurring, proteolytically activated pro-forms of endogenous proteins, is an antibody variant which remains masked for antigen binding until activated locally by an organ-selective protease. This approach is useful to redirect the activity of an antibody recognising a widely distributed target to the site of the underlying pathology of the disease. Pro-antibodies have been described for targeting the tumour microenvironment which has a very specific protease expression signature. My aim is to investigate the use of pro-antibodies for tissue targeting. By using anti-PAI-1 pro-antibodies to specifically target the kidney for the treatment of chronic kidney disease to minimise observed cardiotoxicities, my work aims to validate this technique as a POC. To this end, I have designed a pro-antibody, based on the PAI-1 antibody MEDI-579. Two different types of masking domains are described, peptides and DARPins, which I selected against the MEDI-579 paratope. Using human and rodent transcriptomics data, three proteases, renin, hepsin and MMP-7, were identified that appear restricted to the kidney and have a specific cleavage signature. Activation of the pro-antibody with the renin protease has been demonstrated in vitro and ex vivo and work is on-going to demonstrate the activation in vivo and to characterise the properties of the anti-PAI-1 pro-antibody.","abstract_html":"Antibody-based therapies can have shortcomings if on-target activity causes dose-limiting toxicities. A pro-antibody, inspired by naturally occurring, proteolytically activated pro-forms of endogenous proteins, is an antibody variant which remains masked for antigen binding until activated locally by an organ-selective protease. This approach is useful to redirect the activity of an antibody recognising a widely distributed target to the site of the underlying pathology of the disease. Pro-antibodies have been described for targeting the tumour microenvironment which has a very specific protease expression signature. My aim is to investigate the use of pro-antibodies for tissue targeting. By using anti-PAI-1 pro-antibodies to specifically target the kidney for the treatment of chronic kidney disease to minimise observed cardiotoxicities, my work aims to validate this technique as a POC. To this end, I have designed a pro-antibody, based on the PAI-1 antibody MEDI-579. Two different types of masking domains are described, peptides and DARPins, which I selected against the MEDI-579 paratope. Using human and rodent transcriptomics data, three proteases, renin, hepsin and MMP-7, were identified that appear restricted to the kidney and have a specific cleavage signature. Activation of the pro-antibody with the renin protease has been demonstrated in vitro and ex vivo and work is on-going to demonstrate the activation in vivo and to characterise the properties of the anti-PAI-1 pro-antibody.","abstract_has_math":false,"creators":["Bleuez, Chloe"],"institution":"University of Cambridge","degree_name":"Doctor of Philosophy (PhD)","degree_level":"Doctoral","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Hollfelder, Florian","Jermutus, Lutz"],"committee_chairs":[],"committee_members":[],"year":2021,"date_issued":"2021-09-30","date_published":"2021-09-30","updated_at":"2026-07-22T22:24:17Z","subjects":["antibody","specificity","kidney"],"languages":["eng"],"rights":[],"rights_urls":["https://www.rioxx.net/licenses/all-rights-reserved/"],"identifier_entries":[]},"links":{"outbound_url":"https://doi.org/10.17863/CAM.92119","outbound_label":"DOI","outbound_source":"dc:identifier.doi"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Hollfelder, Florian","Jermutus, Lutz"]},{"key":"dc:contributor.sponsor","label":"Sponsor","values":["Marie-Curie fellowship AstraZeneca"]},{"key":"dc:creator","label":"Author","values":["Bleuez, Chloe"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2021-09-30"]},{"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/1810/344696"]},{"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":["antibody","specificity","kidney"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["https://www.rioxx.net/licenses/all-rights-reserved/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.doi","label":"DOI","values":["10.17863/CAM.92119"]},{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://apollo8-f-pro.lib.cam.ac.uk/bitstreams/333a9634-a9fc-4138-83db-73f82699d923/download"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Antibody-based therapies can have shortcomings if on-target activity causes dose-limiting toxicities. 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