QUANTIFICATION AND ANALYSIS OF BIOMOLECULES VIA MASS SPECTROMETRY FOR THE UNDERSTANDING, DETECTION, AND TREATMENT OF DISEASE

Abstract

Mass spectrometry is an incredibly powerful analytical tool capable of analyzing a wide range of biologically relevant molecules like metabolites and proteins. Metabolites, or molecules with a molecular weight under 1500 daltons, are a group of molecules incredibly sensitive to small changes in the body. This characteristic makes them excellent as diagnostic markers of disease. Metabolites also serve as the building blocks of all of the biochemical pathways that make our body function. By observing changes in metabolism caused by disease, mechanisms of disease and, by extension, potential disease treatments can be discerned. Proteins, a key regulator of metabolism and an important group of molecules for drug development, can also be quantified via mass spectrometry, but their large structure presents several analytical challenges. In this dissertation, the applications of mass spectrometry in the study of metabolites as both a diagnostic biomarker and tool for understanding disease is explored. First, in a mouse model for COVID-19, we used mass spectrometry to identify dysregulation of fatty acids, amino acids, and eicosanoids in the lungs. Furthermore, we observed that dysregulation was worse in peripheral lung tissue despite higher viral loads in central lung tissue. Moving beyond metabolites, chapter 3 of this dissertation focuses on the development of methodology to prepare and analyze proteins using a novel acoustic droplet ejection mass spectrometer. By optimizing instrument and analytical parameters, an analytical pipeline was developed for analyzing a small protein (17 kilodaltons), a medium protein (50 kilodaltons) and a large protein (150 kilodaltons) in under one second per sample. This dissertation provides insight on the detection and mechanism of infectious diseases while also providing insight on new methods to quantify drugs via mass spectrometry.