Detection of DNA Damage: The Synthesis of Aldehyde Reactive Probes for the Quantitation of Abasic Sites

Apurinic/apyrimidinic (AP sites) are the result of damage to DNA. These sites can inhibit replication of the DNA and cause mutations. They are considered to be important intermediates in carcinogenesis and in mutagenesis. It is advantageous to have a simple assay capable of detecting and quantifying these sites in a timely manner. Kubo et al. reported such an assay wherein the aldehyde group in the abasic site reacts with an aldehyde reactive probe (ARP). The number of tagged AP sites is then quantified colorimetrically by an assay similar to an ELISA. The purpose of this research project was to synthesize additional reactive probes that could be used in the detection of abasic sites in DNA. ARP was synthesized using the procedure of Ide etal. Procedures for the synthesis of labeled ARP components, 0-(carboxymethyl)hydroxylamine and biotin hydrazide were developed. Synthetic routes to new ARPs were also investigated. A novel 2-iminobiotin hydrazide hydrochloride probe and a novel hydrazinoacetic acid derivative probe were synthesized and characterized. In addition, a synthetic route to a probe containing a long alkyl tether group was nearly completed.Master of Science in Public Healt