IR-MALDI ION TRAP MASS SPECTROMETER FOR DNA ANALYSIS

DNA分析的质谱学方法具有很高的效率，因为它消除了DNA分析的耗时阶段，即用传统的凝胶电泳和荧光分析方法进行条带识别和解释。我们建议开发一种用于DNA分析的红外(IR)基质辅助激光解吸电离(MALDI)离子陷阱质谱仪(ITMS)，而不是通常用于MALDI离子源的飞行时间(TOF)质谱仪(MS)。在生物分子的MALDI谱中，碎片的存在是非常常见的，这使得即使是简单的DNA混合物的分析也变得非常复杂，特别是当考虑到自动化系统时。这一建议的重点是提高MALDI过程中分子DNA离子的产率，增加方法的灵敏度和可靠性，并允许DNA MS分析的自动化。拟议的ITMS将在几个频率下运行，以将质量范围扩大到m/z 30,000以上。仪器与MALDI离子源的接口将得到优化，以实现更高的灵敏度和更低的碎裂。使用已经在我们的TOFMS实验中测试过的微型可调谐红外激光器，将使我们能够选择最佳的矩阵-波长组合。此外，还将使用冷却的2D芯片样本阵列探头建立一个高通量DNA分析的自动化系统。拟议的商业应用：设想了一些医疗和临床应用，包括核酸(DNA和RNA)分析和诊断、聚合酶链式反应片段分析、蛋白质和多肽分析。还有望用于军事应用，如生物制剂的识别。

The mass spectrometric method of DNA analysis promises to be very productive since it eliminates the time-consuming stages of DNA analysis, namely band identification and interpretation by traditional methods of gel electrophoresis and fluorescent analysis. We propose to develop an infrared (IR) matrix-assisted laser desorption/ ionization (MALDI) ion trap mass spectrometer (ITMS) for DNA analysis in contrast to the time-of-flight (TOF) mass spectrometer (MS) commonly used with MALDI ion sources. The presence of fragments is very common in MALDI spectra of biomolecules, making analysis of even simple DNA mixtures extremely complicated especially when automated systems are considered. This proposal is focused towards improving the yield of the molecular DNA ions in MALDI process, increasing the sensitivity and reliability of the method, and allowing for automation of DNA MS analysis. The proposed ITMS will operate at several frequencies to extend the mass range up to m/z 30,000 and over. The interface of the instrument with the MALDI ion source will be optimized to achieve better sensitivity and lower fragmentation. The use of the miniature tunable infrared laser that has already been tested in the experiments on our TOFMS will allow us to choose the best matrix-wavelength combinations. In addition, an automated system for high throughput DNA analysis will be built using a cooled 2D chip sample array probe. PROPOSED COMMERCIAL APPLICATIONS: A number of medical and clinical applications including Nucleic Acid (DNA & RNA) analysis and diagnostics, PCR fragment analysis, protein and peptide analysis are envisaged. Military applications such as identifications of biological agents are also expected.