LC-IMS-MS Techniques for Human Plasma Profiling

DESCRIPTION (provided by applicant): A New high-throughput, high-coverage proteomics technology that utilizes ion mobility/mass spectrometry (IMS-MS) for the characterization of the plasma proteome will be developed. The IMS-MS instrumentation will be utilized to address the complexity of the plasma proteome by creating a high-dimension, analytical map obtained by analysis of 200 plasma samples. This map will offer a higher resolution of plasma components when compared with commercial technology (by factors of 10 to 50). Work will center on determining the variability in protein abundances as a function of the number of measurements (N) for different individuals across a population as well as for specific individuals as a function of time. The multiple measurements are made possible by a decreased experimental timescale afforded by the new technology. As N increases, the confidence level of protein assignments and abundance determinations also increases. Additionally the ability to perform the requisite N measurements to define "normal" will facilitate the determination of protein abundance change in cardiovascular disease samples (Phase II studies) as what is considered a statistically relevant change will be more accurately known. There are three specific aims to accomplish the work proposed in Phase I studies. These are: 1) Develop LC-IMS-(CID)n-TOF instrumentation as a robust platform for high-throughput plasma proteome analysis; 2) Assess the reproducibility and refine assignments of the "normal" plasma proteome map; and, 3) Initiate comparison of normal" and "diseased" maps for delineation of new biomarkers (primarily carried out in Phase II studies). The construction of a high-coverage proteome map of higher resolution will allow a deeper, broader view of the plasma proteome. Such a vantage point is vital for the determination of protein abundance changes in cardiovascular disease samples (Phase II studies). Observed changes may provide clues to biomarkers associated with cardiovascular disease and may have relevance for diagnostics, therapeutic monitoring, as well as drug discovery studies.