A MALDI-TOF mass spectrometry approach to biomarker discovery and validation

Abstract

Identifying biochemical markers for disease is a difficult endeavor. It requires methods that can quantify multiple components simultaneously, are high throughput, cost effective and deliver high precision and accuracy. In this thesis I explore the potential of matrix-assisted laser desorption/ionization (MALDI) for these applications. The approach developed for this study is a non-targeted MALDI based method to analyze biological tissues and fluids, and here I demonstrate its utility by way of the analysis of human tear fluid. I hypothesize that biomarker signatures of injury, wound repair and/or disease can be detected and quantified in biological fluids by MALDI profiling. I also demonstrate that accurate and precise quantitative results are achievable with coefficients of variation (CV) typically 10% or less, despite the challenges of internal standard selection, sample handling, and ion suppression. In this application, the method offers the ability to detect, identify, and quantify biomarkers associated with eye injury and/or repair and disease. The results of this study reinforce the idea that analysis of human tear fluid can serve as a diagnostic tool, and they illustrate the potential of MALDI-TOF MS for clinical applications.