IR-MALDI ION TRAP MASS SPECTROMETER FOR DNA ANALYSIS

用于 DNA 分析的 IR-MALDI 离子阱质谱仪

• 批准号: 6388322
• 负责人: Vladimir M Doroshenko
• 金额: $ 40.75万
• 依托单位: SCIENCE AND ENGINEERING SERVICES, INC.
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6388322
• 关键词: DNA; biomedical equipment development; electrospray ionization mass spectrometry; high throughput technology; infrared spectrometry; lasers; matrix assisted laser desorption ionization; microarray technology; nucleic acid quantitation /detection; nucleic acid sequence

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.

DNA分析的质谱方法有望非常高效，因为它消除了DNA分析的耗时阶段，即通过凝胶电泳和荧光分析的传统方法进行条带鉴定和解释。 与通常与MALDI离子源一起使用的飞行时间（TOF）质谱仪（MS）相比，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）离子阱质谱仪（ITMS）。 片段的存在在生物分子的MALDI光谱中非常常见，使得即使是简单的DNA混合物的分析也非常复杂，特别是当考虑自动化系统时。 该建议的重点是提高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，以将质量范围扩展到m/z 30，000及以上。 将优化仪器与MALDI离子源的接口，以实现更高的灵敏度和更低的碎片化。已经在我们的飞行时间质谱仪的实验中测试的微型可调谐红外激光器的使用将允许我们选择最佳的矩阵波长组合。此外，将使用冷却的2D芯片样品阵列探针构建用于高通量DNA分析的自动化系统。拟议的商业应用：设想了许多医疗和临床应用，包括核酸（DNA和RNA）分析和诊断、PCR片段分析、蛋白质和肽分析。 预计还可用于军事用途，如识别生物制剂。