Advancement of Multiplexing Strategies to Understand Age and Age-related Disease

Abstract

Proteomics, the large-scale study of proteins and their functions, has furthered understanding of disease and generated new biology hypotheses. Analytical instrumentation such as mass spectrometry is widely utilized in quantitative proteomics to identify hundreds to thousands of proteins and to quantify their relative levels of expression. Multiplexing has increased experimental throughput of quantitative proteomics workflows by enabling the measurement of two to 21 samples in a single analysis and the development of enhanced multiplexing strategies extend multiplexing to more than 50 samples. These strategies result in reduced analysis times, decreased experimental costs, more accurate quantitation, and increased statistical robustness.

In this dissertation, a combined precursor isotopic labeling and isobaric tagging (cPILOT) enhanced multiplexing strategy is improved and implemented to study aging and age-related disease such as Alzheimer’s disease in animal models. Additionally, the development and implementation of quality control considerations for cPILOT and workflow improvements will be discussed.