New proteome techniques: mapping adult D. Melanogaster

Trustees Of Indiana University, Bloomington IN

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Abstract

? DESCRIPTION (provided by applicant): A problem of biomarker discovery efforts is that current analytical methods do not have the capacity to analyze in a timely manner the numerous molecular species within complex biological samples. This problem is exacerbated by the fact that many different samples must be analyzed in order to establish the normal variability across a population for any given molecular species. Here we propose the development of a new analytical instrument that utilizes rapid, gas-phase separations to increase the overall experimental throughput by more than an order of magnitude. The new instrumentation will employ multidimensional ion mobility spectrometry (IMS) techniques to obtain the rapid analysis. By using two dimensions of IMS separations and changing operational parameters such as the electric field and measurement temperature it is anticipated that peak capacities (prior to mass spectrometric analysis) in excess of 104 will be obtained. This is remarkable because it is larger than that obtained from two-dimensional liquid chromatography (LC) separations and requires a fraction of the time. The new instrumentation will be used to characterize the D. melanogaster proteome at different times in the organism lifespan. The significantly increased throughput will allow comparisons of individual organisms with the population mean to better group organisms according to molecular phenotype. Additionally, a novel fluorescent labeling strategy will be employed to better compare the physiological age of individual organisms as opposed to chronological age. The measurement platform will be applied to transgenic Parkinson's and Alzheimer's disease Drosophila in order to determine molecular changes associated with disease progression.