{"id":{"repo_id":"gatech","oai_identifier":"oai:repository.gatech.edu:1853/52149"},"canonical_url":"https://search.dev.ndltd.org/etd/gatech/oai:repository.gatech.edu:1853/52149","repository":{"repo_id":"gatech","name":"Georgia Tech","base_url":"https://repository.gatech.edu/server/oai/request"},"display":{"title":"Reengineering a human-like uricase for the treatment of gout","abstract":"There is an unmet medical need in the treatment of gout. This type of inflammatory arthritis can be efficiently alleviated by the enzyme uricase. This enzyme breaks down uric acid, the causative agent of gout, so it can be flushed from the body. In humans and the other great apes, uricase is a pseudogene and as such is inactive. Research on therapeutic uricases has focused on using enzymes from naturally occurring sources; however, these foreign proteins can be very antigenic and present a potentially life-threatening safety risk to patients. We address the challenges of developing a safer uricase therapeutic by exploiting evidence that, while inactive, the human pseudogene is expressed in the human body and may be recognized as self by the immune system. To develop a ﾓhuman-likeﾔ? uricase we apply the hybrid computational and experimental approach of Ancestral Sequence Reconstruction to search functional sequence space of uricase proteins to engineer an enzyme with high sequence identity to the human pseudogene, and possessing therapeutic levels of activity for the breakdown of uric acid. This dissertation describes the development and characterization of several uricase leads. The most active ancestral uricase possesses both enhanced in vitro and in vivo stability (in healthy rats) when assayed head-to-head Pegloticase, the only FDA approved uricase for the treatment of gout.","abstract_html":"There is an unmet medical need in the treatment of gout. This type of inflammatory arthritis can be efficiently alleviated by the enzyme uricase. This enzyme breaks down uric acid, the causative agent of gout, so it can be flushed from the body. In humans and the other great apes, uricase is a pseudogene and as such is inactive. Research on therapeutic uricases has focused on using enzymes from naturally occurring sources; however, these foreign proteins can be very antigenic and present a potentially life-threatening safety risk to patients. We address the challenges of developing a safer uricase therapeutic by exploiting evidence that, while inactive, the human pseudogene is expressed in the human body and may be recognized as self by the immune system. To develop a ﾓhuman-likeﾔ? uricase we apply the hybrid computational and experimental approach of Ancestral Sequence Reconstruction to search functional sequence space of uricase proteins to engineer an enzyme with high sequence identity to the human pseudogene, and possessing therapeutic levels of activity for the breakdown of uric acid. This dissertation describes the development and characterization of several uricase leads. The most active ancestral uricase possesses both enhanced in vitro and in vivo stability (in healthy rats) when assayed head-to-head Pegloticase, the only FDA approved uricase for the treatment of gout.","abstract_has_math":false,"creators":["Kratzer, James Timothy"],"institution":"Georgia Institute of Technology","degree_name":null,"degree_level":"Doctoral","degree_discipline":null,"degree_department":"Chemistry and Biochemistry","school":null,"contributors":[],"advisors":["Gaucher, Eric A."],"committee_chairs":[],"committee_members":["Hud, Nick V.","Kelly, Wendy L.","Williams, Loren D.","Ortlund, Eric A."],"year":2013,"date_issued":"2013-04-29","date_published":"2013-04-29","updated_at":"2026-07-27T19:49:22Z","subjects":["Gout","Uricase","Ancestral sequence reconstruction","Protein engineering","Evolutionary synthetic biology","Pseudogene","Enzyme replacement","Therapy","Enzyme assays","Protein purification","Pharmacokinetics","Protein solubility","Evolution"],"languages":["en_US"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/1853/52149","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Gaucher, Eric A."]},{"key":"dc:contributor.committeemember","label":"Committee Member","values":["Hud, Nick V.","Kelly, Wendy L.","Williams, Loren D.","Ortlund, Eric A."]},{"key":"dc:contributor.department","label":"Department","values":["Chemistry and Biochemistry"]},{"key":"dc:creator","label":"Author","values":["Kratzer, James Timothy"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2014-08-27T13:31:08Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2014-08-28T05:30:04Z"]},{"key":"dc:date.issued","label":"Date","values":["2013-04-29"]},{"key":"dc:publisher","label":"Institution","values":["Georgia Institute of Technology"]},{"key":"dc:type","label":"Dc Type","values":["Text"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Doctoral"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Gout","Uricase","Ancestral sequence reconstruction","Protein engineering","Evolutionary synthetic biology","Pseudogene","Enzyme replacement","Therapy","Enzyme assays","Protein purification","Pharmacokinetics","Protein solubility","Evolution"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en_US"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/1853/52149"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["There is an unmet medical need in the treatment of gout. This type of inflammatory arthritis can be efficiently alleviated by the enzyme uricase. This enzyme breaks down uric acid, the causative agent of gout, so it can be flushed from the body. In humans and the other great apes, uricase is a pseudogene and as such is inactive. Research on therapeutic uricases has focused on using enzymes from naturally occurring sources; however, these foreign proteins can be very antigenic and present a potentially life-threatening safety risk to patients. We address the challenges of developing a safer uricase therapeutic by exploiting evidence that, while inactive, the human pseudogene is expressed in the human body and may be recognized as self by the immune system. To develop a ﾓhuman-likeﾔ? uricase we apply the hybrid computational and experimental approach of Ancestral Sequence Reconstruction to search functional sequence space of uricase proteins to engineer an enzyme with high sequence identity to the human pseudogene, and possessing therapeutic levels of activity for the breakdown of uric acid. This dissertation describes the development and characterization of several uricase leads. The most active ancestral uricase possesses both enhanced in vitro and in vivo stability (in healthy rats) when assayed head-to-head Pegloticase, the only FDA approved uricase for the treatment of gout."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["Ph.D."]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Reengineering a human-like uricase for the treatment of gout"]}]}],"canonical_facts":{"dc:contributor.advisor":["Gaucher, Eric A."],"dc:contributor.committeemember":["Hud, Nick V.","Kelly, Wendy L.","Williams, Loren D.","Ortlund, Eric A."],"dc:contributor.department":["Chemistry and Biochemistry"],"dc:creator":["Kratzer, James Timothy"],"dc:date.accessioned":["2014-08-27T13:31:08Z"],"dc:date.available":["2014-08-28T05:30:04Z"],"dc:date.issued":["2013-04-29"],"dc:description.abstract":["There is an unmet medical need in the treatment of gout. This type of inflammatory arthritis can be efficiently alleviated by the enzyme uricase. This enzyme breaks down uric acid, the causative agent of gout, so it can be flushed from the body. In humans and the other great apes, uricase is a pseudogene and as such is inactive. Research on therapeutic uricases has focused on using enzymes from naturally occurring sources; however, these foreign proteins can be very antigenic and present a potentially life-threatening safety risk to patients. We address the challenges of developing a safer uricase therapeutic by exploiting evidence that, while inactive, the human pseudogene is expressed in the human body and may be recognized as self by the immune system. To develop a ﾓhuman-likeﾔ? uricase we apply the hybrid computational and experimental approach of Ancestral Sequence Reconstruction to search functional sequence space of uricase proteins to engineer an enzyme with high sequence identity to the human pseudogene, and possessing therapeutic levels of activity for the breakdown of uric acid. This dissertation describes the development and characterization of several uricase leads. The most active ancestral uricase possesses both enhanced in vitro and in vivo stability (in healthy rats) when assayed head-to-head Pegloticase, the only FDA approved uricase for the treatment of gout."],"dc:description.degree":["Ph.D."],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["http://hdl.handle.net/1853/52149"],"dc:language.iso":["en_US"],"dc:publisher":["Georgia Institute of Technology"],"dc:subject":["Gout","Uricase","Ancestral sequence reconstruction","Protein engineering","Evolutionary synthetic biology","Pseudogene","Enzyme replacement","Therapy","Enzyme assays","Protein purification","Pharmacokinetics","Protein solubility","Evolution"],"dc:title":["Reengineering a human-like uricase for the treatment of gout"],"dc:type":["Text"],"thesis:degree_level":["Doctoral"]},"updated_at":"2026-07-27T19:49:22Z"}