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

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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.