Development of Advanced Time-of-flight (TOF)-Mass Spectrometer(MS) for Proteomics

大卫·H博士获得了一笔赠款。Russell与德克萨斯A M大学（TAMU）合作，开发基质辅助激光解吸电离（MALDI）高分辨率飞行时间（TOF）质谱，用于快速、高灵敏度的肽和蛋白质分析。 仪器开发项目是TAMU科学家和三家领先的质谱仪器制造商的合作。 项目目标的成功完成将为大分子质谱提供即时改进。 拟议研究的第二个目标涉及开发一种高灵敏度、周期性聚焦离子迁移率（IM）漂移管，该漂移管将用于快速分离复杂生物混合物中存在的肽和/或蛋白质。 随着IM漂移管的研制成功，它将被用于高分辨率TOF仪器。 IM和高分辨率TOF质谱的组合将为蛋白质鉴定提供增强的能力，特别是蛋白质表达、翻译后修饰和肽/蛋白质测序的研究。 高分辨率TOF质谱仪的实际构建涉及对最初由ABI-PerSeptive，Inc.设计和构建的现有仪器的广泛修改。并于1994年安装在TAMU。 该仪器是非常独特的，只有3个这样的仪器曾经建成，和基本的仪器平台仍然代表了TOF质量分辨率（15，000 - 25，000的质量分辨率是常规）和质量测量精度（质量误差为10 ppm）的尖端能力。 仪器的性能将通过增加新的电子设备和专门设计的多阳极离子探测器来提高，反射器的几何形状将从原来的共线配置改为1.5度反射角，以改善离子传输和时间聚焦。 预计升级后的仪器质量分辨能力为75，000 - 100，000，质量测量精度为0.5-3 ppm。 周期聚焦离子迁移率漂移管（由TAMU申请专利）将在TAMU设计和制造，并由工业合作者改装为高分辨率TOF仪器。 在过去的十年中，质谱已经发展成为生物学和化学生物学中最广泛使用的分析方法之一。 在现代质谱仪中，TOF仪器是用户最友好和功能最多的，并且TOF仪器被生物学家和化学生物学家最广泛地使用。 因此，新的高性能TOF仪器的开发将广泛影响生物学研究。 PI已经开发了大量的生物学和化学生物学合作者，这确保了拟议的仪器将被TAMU和全国各地的合作者有效地使用。 生物学家和化学生物学家面临的一个主要挑战涉及表征和识别少量存在的生物材料是高度复杂的混合物，IM和高分辨率TOF仪器的发展具有很大的潜力，以提高高分子质谱的性能标准。 仪器的发展和伴随的方法的发展将在未来的生物分析化学家和化学生物学家的培训产生重大影响。 最后，拟议的研究符合MRI计划的目标，旨在促进学术研究人员和私营部门仪器开发商之间的伙伴关系。

A grant has been awarded to Dr. David H. Russell at Texas A&M University (TAMU) to develop matrix-assisted laser desorption ionization (MALDI) high resolution time-of-flight (TOF) mass specroscopy for rapid, high-sensitivity analysis of peptides and proteins. The instrument development project is a collaboration involving scientist at TAMU and three of the leading manufacturers of mass spectrometry instruments. Successful completion of the project objectives will provide immediate improvements for macromolecule mass spectrometry. A secondary objective of the proposed research involves the development of a high-sensitivity, periodic focusing ion mobility (IM) drift tube, which will be used for rapid separation of peptides and/or proteins present in complex biological mixtures. Following successful development of the IM drift tube it will be adapted to the high resolution TOF instrument. The combination of IM and high resolution TOF mass spectrometry will provide enhanced capabilities for protein identification, especially studies of protein expression, post-translational modifications and peptide/protein sequencing. The actual construction of the high resolution TOF mass spectrometer involves extensive modifications to an existing instrument initially designed and constructed by ABI-PerSeptive, Inc. and installed at TAMU in 1994. The instrument is highly unique, only 3 such instruments were ever built, and the basic instrument platform still represents cutting-edge capabilities in TOF mass resolution (mass resolving powers of 15,000-25,000 are routine) and mass measurement accuracy (mass errors of 10 ppm). The instrument performance will be enhanced by addition of new electronics and specially designed multi-anode ion detectors, and the geometry of the reflectron will be changed from the original co-linear configuration to a 1.5 degree reflecting angle to improve ion transmission and temporal focusing. Mass resolving power of 75,000-100,000 and mass measurement accuracy of 0.5-3 ppm is projected for the up-graded instrument. The periodic focusing ion mobility drift tube (patented by TAMU) will be designed and constructed at TAMU, and adapted to the high resolution TOF instrument by the industrial collaborators. In the last decade mass spectrometry has evolved as one of the most widely used analytical methods in biology and chemical-biology. Of the modern mass spectrometers, TOF instruments are the most user-friendly and versatile, and TOF instruments are the most widely used by biologist and chemical-biologist. Consequently, development of new, high performance TOF instruments will broadly impact biological research. The PI has developed a large number of biology and chemical-biology collaborators, which insures that the proposed instrument will be effectively used by collaborators at TAMU and across the nation. A major challenge facing biologist and chemical-biologist involves characterization and identification of small amounts of biological materials present is highly complex mixtures, and the development of IM and high-resolution TOF instruments have great potential to raise the performance bar for macromolecule mass spectrometry. The development of the instrumentation and the concomitant methods development will have significant impact in the future training of bio-analytical chemists and chemical-biologists. Lastly, the proposed research meets goals of the MRI program aimed at promoting partnerships between academic researchers and private sector instrument developers.