Bioanalytical Approaches to Chemical Biology
Harry, Stephen Reese
As the field of chemical biology evolves, so does the development of novel bioanalytical techniques capable of accurately measuring biochemical composition and regulation. This dissertation explores three aspects of chemical biology using validated analytical methodology coupled with novel nanoscale intracellular detection probes. Using LC tandem mass spectrometry, identification and quantification of hydroxylated fatty acids associated with S. mansoni hemozoin was achieved. Additionally, the metabolic impact of 4-hydroxynonenal toxicity on macrophage activation was examined using multianalyte microphysiometry which confirmed a concentration and temporal inhibition of overall cellular metabolism. Utilizing the inherent characteristics of DNA-coated gold nanospheres, a new mRNA detection probe was developed that positively identifies tyrosinase-expressing melanoma cells. The broad scope of this work explores the use of new and emerging bioanalytical techniques and reagents that will be requisite for understanding biochemical activity at the single-cell level.
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