While the catalytically powerful [Cu,Zn] superoxide dismutase (SOD1) possesses great potential as a therapeutic, unfavorable properties in circulation limit its use in clinical medicine. The small, water soluble dimer is rapidly excreted by the kidney. Previous initiatives have been used to increase the mass of the enzyme (PEGylation, liposome encapsulation). This has resulted in highly heterogeneous mixtures of modified SOD1, which are difficult to characterize. Furthermore, these modified proteins have utilized foreign material that has shown to elicit an inflammatory response. We developed an improved strategy that creates a homogenous high molecular weight SOD1 based on combinations of the protein itself. This was accomplished through the addition of a site-specific, azide functionalized cross-linker to unmodified SOD1, followed by the conjugation of SOD dimers using CuAAC and a bis-alkyne linker to form a 64 kDa SOD tetramer. The final product, bis-SOD, presents the fully catalytic activity of the combined proteins.
| Identifer | oai:union.ndltd.org:TORONTO/oai:tspace.library.utoronto.ca:1807/42934 |
| Date | 28 November 2013 |
| Creators | Siren, Erika |
| Contributors | Kluger, Ronald |
| Source Sets | University of Toronto |
| Language | en_ca |
| Detected Language | English |
| Type | Thesis |
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