New proteome techniques: mapping adult D. Melanogaster

 描述(申请人提供)：生物标记物发现工作的一个问题是，目前的分析方法不能及时分析复杂生物样品中的众多分子物种。必须对许多不同的样本进行分析，以确定任何给定分子物种在整个种群中的正常可变性，这一事实加剧了这个问题。在这里，我们建议开发一种新的分析仪器，它利用快速的气相分离来增加总体实验吞吐量一个数量级以上。新的仪器将使用多维离子迁移率光谱(IMS)技术来获得快速分析。通过使用两个维度的IMS分离和改变操作参数，如电场和测量温度，预计将获得超过104的峰容量(在质谱分析之前)。这是值得注意的，因为它比二维液相色谱(LC)分离得到的更大，并且只需要一小部分时间。这种新的仪器将被用来描述生物体生命周期中不同时间的黑腹果蝇蛋白质组。显著增加的吞吐量将允许将单个生物体与种群平均值进行比较，以便根据分子表型更好地将生物体分组。此外，一种新的荧光标记策略将被用来更好地比较单个生物体的生理年龄，而不是时间年龄。该测量平台将应用于转基因帕金森氏症和阿尔茨海默病果蝇，以确定与疾病进展相关的分子变化。

 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.