The development of analytical techniques for the detection and characterization of biomolecules

Abstract

This dissertation presents diverse projects that all deal with the developments of analytical methods for the detection and characterization of biomolecules. More specifically, the two chapters following the introduction will discuss methods developed for the detection of biomarkers that have been implicated in various types of cancer. Chapter 1 provides an overview of the analytical techniques and instrumentation used to produce data presented in this thesis. Emphasis is placed on the mass spectrometric and differential mobility instrumentation used for the detection of biomolecules. Chapter 2 presents a high throughput method for the detection of dG-ABP by differential mobility-mass spectrometry. It capitalized on the abilities of DMS to rapidly filter normal nucleosides, enzymes and buffers in less than a second to replace traditional chromatographic methods that require times up to 1.5 hours. The developed procedures were able to detect adducts in DNA extracted from a human cell line treated with dG-ABP. This work was presented at the 55th ASMS Conference on Mass Spectrometry, Indianapolis, IN, June 3-7, 2007. Chapter 3 describes methods developed with commercially available instrumentation for the development of LC-MS/MS methods in order to detect and characterize the phosphorylation state of Tpl2. The systematic method described was able to detect two novel sites of phosphorylation. This work was published in The Journal of Proteome Research (2007). Chapter 4 describes the development of a forensic chemistry course for chemistry majors. It will include a discussion of the process of writing the course as well as student feedback offered at the completion of the course.