Development of Next Generation Mass Spectrometric Instrumentation for Proteomics

项目摘要/摘要 目前，蛋白质组学的大多数质谱学(MS)分析都是按顺序进行的 模式，在这种模式下，一个样品中的各种物种被一个接一个地选择和询问。AS 依次检查每个物种所需的有限时间的结果，顺序模式MS 在灵敏度、速度和分析所有离子的能力方面都受到不可避免的限制，尤其是 当离子束的成分复杂且快速变化时。这些限制具有 保存了大量的生物学和生物医学，例如，包括深层单细胞蛋白质组 分析，目前的MS技术是遥不可及的。在本提案中，我们假定 通过并行执行MS可以大大提高灵敏度、速度和动态范围(通过 类似于下一代DNA测序)，从而克服了 目前商用的质谱计主要以顺序模式运行。 在这里，我们建议开发新的MS工具来以大规模并行的方式执行MS 态度，考虑到两个主要目标： 目标1.提高蛋白质组分析的灵敏度、速度和深度 最终超过1000倍。当前的顺序MS方法可以比作采样 尼亚加拉瀑布有一个水桶，大部分样本都被浪费了。我们建议的平行 MS技术旨在消除这种巨大的浪费。 目的2.在MS分析之前实时滤除噪声以消除不需要的离子背景， 从而最大化了感兴趣的样品离子的MS利用率和所产生的信号比。 噪声比，以及提供更大的动态范围来测量非常低的丰度 在存在高度丰富的成分的情况下。 成功实现这些目标将允许深入、全面、高吞吐量 在样本可获得性有限的情况下对蛋白质组进行分析，其中 由于其丰度低，或当需要进行单细胞分析时，感兴趣的细胞无法检测到 解决手头的生物学或生物医学问题。这一努力的成功将推动 提出了许多需要蛋白质组学分析的基础和应用生物医学研究领域 类似于通过开发Next所取得的巨大进步 世代DNA测序。

Project Summary/Abstract Currently, most mass spectrometric (MS) analyses for proteomics are performed in sequential mode, wherein various species in a sample are selected and interrogated one after another. As a consequence of the finite time needed to examine each species in turn, sequential mode MS suffers from inescapable limitations in sensitivity, speed and ability to analyze all ions, especially when the composition of the ion beam is complex and rapidly changing. These limitations have kept vast tracts of biology and biomedicine, including, for example, deep single cell proteome analysis, out of reach of the current MS technology. In the present proposal, we posit that sensitivity, speed and dynamic range can be vastly improved by performing MS in parallel (by analogy to Next Generation DNA Sequencing), thus overcoming the technical barriers inherent to current commercial mass spectrometers that operate largely in sequential mode. Here, we propose to develop new MS instrumentation to execute MS in a massively parallel manner, with two major objectives in mind: Objective 1. Increase the sensitivity, speed and depth of proteome analyses by up to and ultimately beyond 1000-fold. The current sequential MS approaches can be likened to sampling the Niagara Falls with a bucket, where the majority of the sample is wasted. Our proposed parallel MS technology is designed to eliminate this immense waste. Objective 2. Filter noise in real time to eliminate unwanted ion background prior to MS analysis, thereby maximizing the MS utilization of the sample ions of interest and the resulting signal-to- noise ratios, as well as providing increased dynamic range to measure very low abundance components in the presence of highly abundant components. Successful attainment of these objectives will allow deep, comprehensive, high throughput analyses of proteomes in cases where sample availability is limiting, where the components of interest elude detection due to their low abundance, or when single cell analysis is needed to address the biological or biomedical question at hand. Success in this endeavor will propel forward many areas of basic and applied biomedical research that require proteomic analyses in a manner analogous to the immense progress that has been made through development of Next Generation DNA Sequencing.