A High-Repetition-Rate IR Laser for Mass Spectrometry

描述(由申请人提供)：该第二阶段SBIR研究计划的目标是继续开发用于高通量和高灵敏度红外MALDI应用的高重复频率可调红外激光系统。该计划的目标是产生一个紧凑的原型激光系统，并展示新的高重复率IR-MALDI应用。该项目的具体计划任务是：开发和测试基于商用泵浦激光系统的第二代紧凑型高重复频率OPO；在MALDI飞行时间(TOF)质谱仪上演示第二代OPO；将第二代OPO与正交提取MALDI离子源混合四极飞行时间质谱仪进行接口和演示；以及开发用于空间分辨凝胶和组织分析的扫描红外MALDI和LDI质谱仪。 传统的MALDI使用低重复频率激光系统，当必须分析大量样品时，或在印迹、直接凝胶或组织分析等尖端扫描/测绘应用中，测量时间可能会很长。减少分析时间对于包括功能蛋白质组学和药物发现在内的几个领域的商业大容量实验室非常重要。最近的研究集中在利用高重复频率(1 KHz)的UV激光系统来提高UV-MALDI和AP-MALDI实验的吞吐量和灵敏度。然而，由于缺乏合适的激光系统，高重复频率红外MALDI与UV MALDI相比具有许多优点，包括分析大质量DNA和其他高分子生物分子，分析凝胶斑点或直接从细胞或组织样品中分析生物聚合物，以及分析药物和其他小分子分子(200-1500 Da)。 该计划将通过开发和测试小型激光原型系统，并展示其在一些关键应用中的能力，满足MALDI应用对高重复频率中红外激光系统的需求，这些应用将受益于数据采集率和离子通量的增加。 生物分子的质谱学是一项关键的生物分析技术，它直接作为临床诊断的组成部分和生化研究的重要工具在公共卫生中发挥着重要作用，有助于我们更好地了解蛋白质和基因的基本生化过程以及它们与环境之间的相互关系。MALDI质谱学等关键技术提供了丰富的蛋白质组学数据，极大地提高了我们对各种蛋白质及其在生命系统中的分布和功能的了解。拟议的工作将通过提供一种能够提高数据收集速度和各种应用中的数据质量的高重复频率红外激光器来进一步提高MALDI的性能。

DESCRIPTION (provided by applicant): The goal of this Phase II SBIR research program is to continue development of a high-repetition-rate tunable IR laser system for high-throughput and high-sensitivity IR MALDI applications. The program goals are to generate a compact prototype laser system and to demonstrate novel high repetition-rate IR-MALDI applications. The specific program tasks of the project are: Develop and test the second-generation compact high repetition-rate OPO based on a commercially available pump laser system; Demonstrate the second-generation OPO on a MALDI time-of-flight (TOF) mass spectrometer; Interface and demonstrate the second-generation OPO with an orthogonal extraction MALDI ion source hybrid quadrupole-time-of-flight mass spectrometer; and develop scanning IR MALDI and LDI mass spectrometry for spatially resolved gel and tissue analysis. Traditional MALDI has utilized low-repetition-rate laser systems, which can yield lengthy measurement times when large numbers of samples must be analyzed, or in cutting-edge scanning/mapping applications such as blot, direct gel or tissue analysis. Reducing analysis times is important for commercial high-volume laboratories in several areas including functional proteomics and drug discovery. Recent research has focused on utilizing high-repetition-rate (1 kHz) UV laser systems for both improved throughput and increased sensitivity in UV-MALDI and AP-MALDI experiments. However due to lack of suitable laser systems, there has been no research in the area of high repetition- rate IR MALDI, which has several advantages over UV MALDI including analysis of high-mass DNA and other high molecular weight biomolecules, analysis of biopolymers from gel blots or directly from cell or tissue samples, and the analysis of pharmaceuticals and other low molecular weight molecules (200 - 1500 Da). This program will address the need for a high-repetition rate mid-IR laser system for MALDI applications by developing and testing a prototype compact laser system, and demonstrating its capabilities in a number of key applications which will benefit from the increase in data collection rate and ion flux. Mass spectrometry of biological molecules is a key bioanalytical technology, and plays an important part in public health both directly as a component of clinical diagnostics and as a key tool in biochemical research, enhancing our understanding of the basic biochemical processes of proteins and genes and their interrelation among their environment. Key techniques such as MALDI mass spectrometry have provided a wealth of proteomics data which has greatly enhanced our knowledge of the various proteins and their distribution and function in living systems. The proposed work will further enhance MALDI performance by providing a high-repletion-rate IR laser capable of improving both the speed of data collection but also the quality of the data in a variety of applications.