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IR MALDI应用的高重复率可调谐IR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：基于商用泵浦激光系统开发和测试第二代紧凑型高重复率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离子源混合四极杆飞行时间质谱仪进行接口和演示;并开发扫描IR MALDI和LDI质谱法，用于空间分辨凝胶和组织分析。 传统的MALDI使用低重复率激光系统，当必须分析大量样品时，或在尖端扫描/绘图应用中，如印迹、直接凝胶或组织分析，这可能会产生较长的测量时间。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多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）的紫外激光系统，提高吞吐量和灵敏度的UV-MALDI和AP-MALDI实验。然而，由于缺乏合适的激光系统，在高重复率IR MALDI领域没有研究，其具有优于UV MALDI的几个优点，包括分析高质量DNA和其它高分子量生物分子，分析来自凝胶印迹或直接来自细胞或组织样品的生物聚合物，以及分析药物和其它低分子量分子（200 - 1500 Da）。 该计划将通过开发和测试原型紧凑型激光系统来解决MALDI应用对高重复率中红外激光系统的需求，并展示其在一些关键应用中的能力，这些应用将受益于数据收集速率和离子通量的增加。 生物分子的质谱分析是一项关键的生物分析技术，在公共卫生中发挥着重要作用，无论是直接作为临床诊断的组成部分，还是作为生物化学研究的关键工具，增强了我们对蛋白质和基因的基本生物化学过程及其与环境之间相互关系的理解。MALDI质谱等关键技术提供了丰富的蛋白质组学数据，极大地增强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。拟议的工作将进一步提高MALDI的性能，提供一个高重复率红外激光器，能够提高数据收集的速度，但也在各种应用中的数据质量。

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.