Masters Thesis

Isolation of aptamers for galactonate and synthesis of cryptophane derivative precursors

Galactonate-binding aptamers were isolated using a technique called Systematic Evolution of Ligands by Exponential Enrichment (SELEX). This iterative process was able to remove unwanted aptamers from a pool of oligonucleotide library. The target molecule (galactonate-Shinkai complex) was eventually reduced to 250 and 10 μM after several rounds of SELEX and observing selectivity. During counter-selection however, it showed that the aptamers were heavily binding to 100 μM Shinkai. Additional SELEX rounds were applied to further enrich the DNA pool. Insertion of the isolated DNAs in E. coli did not provide enough bacterial colonies for DNA sequencing. A 20-step synthetic route was proposed to synthesize two cryptophane molecules. The cryptophane-cage molecule is able to bind with 129Xe which is used in contrast agents to improve signal detection. Precursors for a cryptophane cage molecule were successfully synthesized and characterized. Additional works are needed to complete the proposed steps.

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