High Resolution MALDI-TOF Mass Spectrometer

描述(申请人提供)：用于多肽和小分子的高性能MALDI-TOF MS摘要。该项目将以MALDI-TOF质谱学的最新进展为基础，开发新的仪器和软件，提供与FTICR和Orbitrap仪器上的电喷雾电离相媲美的分辨率和质量精度，但速度、吞吐量、灵敏度和动态范围都比任何电喷雾系统都大几个数量级。在初步工作中，已经开发出一种原型仪器，在选定的质量上提供超过30,000的分辨率，并在已知蛋白质消化的大质量范围内提供ppm质量精度。根据初步结果和理论计算，在小型、廉价的台式仪器中，最终分辨率预计至少为100,000，大幅面MALDI板上所有斑点的质量精度优于1ppm RMS质量精度，使用自动内部校准，无需操作员干预和灵敏度，以检测和准确确定1阿托摩尔水平的质量。该项目的长期目标是提供实用的商业仪器，使高性能MALDI-TOF质谱学的全部功率常规可用于目前可用的工具无法令人满意的困难生物应用。这项工作将包括飞行时间质谱仪的技术进步，还将侧重于具体应用的需要，以及开发满足这些需要的可靠和具有成本效益的手段。公共卫生相关性：拟议的仪器是基于MALDI的新的综合分析系统的重要组成部分，该系统将使大规模蛋白质组学和代谢组学变得实用。人们普遍预测，人类基因组计划的完成以及复杂生物样本中蛋白质和小分子分析方法的改进，将在不久的将来极大地改变药物发现和医学临床实践，但这尚未实现。目前的技术显然不能胜任这项任务。

DESCRIPTION (provided by applicant): High-performance MALDI-TOF MS for Peptides and Small Molecules Summary. This project will build on recent advances in MALDI-TOF mass spectrometry to develop new instruments and software that provide resolving power and mass accuracy competitive with electrospray ionization on FTICR and Orbitrap instruments, but with speed, throughput, sensitivity and dynamic range orders of magnitude greater than are possible with any electrospray system. In preliminary work a prototype instrument has been developed that provides greater than 30,000 resolving power at a selected mass, and ppm mass accuracy over a broad mass range on known protein digests. Based on preliminary results and theoretical calculations the ultimate resolving power is expected to be at least 100,000 in a small, inexpensive benchtop instrument with better than 1ppm RMS mass accuracy for all spots on a large format MALDI plate using automated internal calibration with no operator intervention and sensitivity to detect and accurately determine masses at the 1 attomole level. The long-range goal of this project is to provide practical commercial instruments that make the full power of high-performance MALDI-TOF mass spectrometry routinely available for difficult biological applications that are not satisfactorily served by the tools presently available. This effort will include technical advances in TOF MS and will also focus on the needs of specific applications and on developing reliable and cost-effective means for satisfying these needs. PUBLIC HEALTH RELEVANCE: The proposed instrument is a vital component of new integrated analytical systems based on MALDI that will make large-scale proteomics and metabolomics practical. Completion of the human genome project and improved methods for analyzing proteins and small molecules in complex biological samples have been widely predicted to dramatically change drug discovery and clinical practice of medicine in the near future, but this has not yet been realized. Current technology is clearly not up to the task.