dc.contributor.advisor | Tolbert, Thomas J | |
dc.contributor.author | Hutchison, Kevin Michael | |
dc.date.accessioned | 2016-10-11T19:16:56Z | |
dc.date.available | 2016-10-11T19:16:56Z | |
dc.date.issued | 2015-12-31 | |
dc.date.submitted | 2015 | |
dc.identifier.other | http://dissertations.umi.com/ku:14300 | |
dc.identifier.uri | http://hdl.handle.net/1808/21664 | |
dc.description.abstract | Interleukin 1 (IL-1) is an inflammatory cytokine that helps the immune system fight disease. The inflammatory action of IL-1 is regulated by the naturally occurring interleukin 1 receptor antagonist (IL-1ra). IL-1ra contains one N-linked glycosylation site and is expressed in humans in both glycosylated and non-glycosylated forms. The one current IL-1ra drug available (Kineret®) is expressed in Escherichia coli as a non-glycosylated form. Because of this, most studies of IL-1ra have been done with the non-glycosylated form and the effects of glycosylation have not been studied. This research attempts to study the effects of the glycosylation of IL-1ra with its binding to the interleukin 1 receptor type 1 (IL-1R1). Both IL-1R1 and IL-1ra were expressed in a glycosylation deficient strain of Pichia pastoris. In-vitro binding experiments between the two proteins were studied using both biolayer interferometry and size exclusion chromatography. Due to the long dissociation rate between IL-1ra and IL-1R1, biolayer interferometry was an unfeasible method. Size exclusion chromatography, however, proved to be a promising tool to study binding. Initial experiments with non-glycosylated IL-1ra showed a potential KD of 1.7 nM. Future studies need to be done using the glycosylated form of IL-1ra to determine any differences in binding between the glycosylated form and the non-glycosylated form. | |
dc.format.extent | 59 pages | |
dc.language.iso | en | |
dc.publisher | University of Kansas | |
dc.rights | Copyright held by the author. | |
dc.subject | Pharmaceutical sciences | |
dc.title | Role of Glycosylation of IL-1ra on its Binding to IL-1R1 | |
dc.type | Thesis | |
dc.contributor.cmtemember | Siahaan, Teruna J | |
dc.contributor.cmtemember | Wang, Zhuo | |
dc.thesis.degreeDiscipline | Pharmaceutical Chemistry | |
dc.thesis.degreeLevel | M.S. | |
dc.provenance | 04/05/2017: The ETD release form is attached to this record as a license file. | |
dc.rights.accessrights | openAccess | |