Is a NOESY-like 2 dimensional experiment possible in mass spectrometry?

This research is intended to develop better instrumentation which will be helpful in a wide variety of chemical and biochemical problems, such as understanding the fundamental mechanisms of disease and the migration of petroleum byproducts through ground water.

For example, type 1 diabetes is a condition where some event has triggered the body's immune system to kill the islet cells in the pancreas that produce insulin. The exact molecular change that this virus produces to trigger this immune response is also not clear, but it usually stimulates antibody production against insulin itself, insulin precursor protein, or islet cell surface proteins such as GAD65. In order to understand exactly what the problem is, and hence to be able to develop more effective treatment and hopefully a cure, it is important to figure out the nature of that exact change. Unfortunately, this is a classic "needle in a haystack" problem. There are tens of thousands of proteins in most tissues, and although the medical/scientific community has a number of good guesses about the proteins that are involved and the modifications that are made to them, the sample is nevertheless very complex. It is possible to separate most of the components of these mixtures, but even when the candidate molecules are isolated, it is necessary to sequence them completely, without disrupting the modifications in order to find out exactly what changes. Mass Spectrometry methods exist to do this, but they all have limitations. This research progress seeks to develop a new, unbiased method to completely fragment proteins and other biomolecules in complex mixtures without pre-selecting which ones to focus on. By simultaneously generating fragment mass information on all components in the mixture, more data can be generated, faster, and with little bias which can then be data-mined for the important information.

Another example involves tracing of potential sources of contamination of ground water (and fish) by petroleum byproducts from refineries and testing of bioremediation methods. The phasing control methods to be developed herein will improve our ability to determine the presence and structure of oil-related compounds in many samples, from soil, to ground-water, to bioremediation algae.

The primary method involved, called two-dimensional Fourier Transform (2DFT) mass spectrometry is a corollary of Nuclear Magnetic Resonance correlation spectroscopy methods such as NOESY. The 2DFT method has actually been around since the late 1980's, but it was impractical at that time. That impracticality was a side-effect of the fragmentation methods used at the time, but can be avoided by use of new methods which have become available, and even widespread, in the last decade. Furthermore, this methodology can potentially be applied to all sorts of ions without much tinkering with the controlling voltages, currents, and laser power. If this promise bears fruit, new instruments can be designed and built which will make this type of analysis routinely available to the bulk of biomedical researchers.

This method is, of course, not limited to the study of diabetes and petroleum contamination studies, but can be used in the study of any modification on any molecule, whether it's related to heart disease or anthrax vaccines. Thus, it will be of great use in the pharmaceutical, chemical, petroleum, and biotech industries in terms of development, quality control, tracing, and troubleshooting. It can, theoretically, be applied to any complex mixture, provided the fragmentation is effective for such molecules. As always, with science, the devil is in the details, and the methods being developed in this project will generate far more detailed information on complex samples than previously possible.

This research involves development of new methods to extend the data density in tandem mass spectrometry experiments by designing 2D methodologies analogous to NOESY experiments in NMR. In doing so, it offers the ability to generate MS/MS data on all components in a mixture in an unbiased, automatic method where the fragment ion abundances are programmatically modulated to link precursors with products. This has advantages over traditional MS/MS methods in that no precursor ion selection (and the associated experimental biases that introduces) is required. Ultimately, such basic science developments that increase the performance of mass spectrometers will extend far and wide and may, for example, allow detection of the exact modifications that trigger type 1 diabetes. Such fundamental instrumental developments cannot be overestimated; for example, Gallileo was able to revolutionize our understanding of the universe simply because he was able to develop his own, improved telescopes. Most progress in science can be directly attributed to improved instrumentation which allow the researcher to see farther, with higher resolution/accuracy/sensitivity, etc.

PhD students trained in advanced mass spectrometry, analytical science, complex mixture analysis, and the chemistry in the petroleum industry will be very well placed for a strong future in industry and/or academia. In order to make certain of this, PBO has historically maintained a high publication rate for each PhD thesis with the students being trained to communicate their results in publication form, which is then also included in their theses. When combined with conference presentations, these students are very employable. This is another direct economic benefit of this research.

To increase impact, this project will be particularly highlighted during the 10th European FTMS workshop to be organized by our research group at Warwick in April, 2012. This workshop, the ideal venue for this research, will be coordinated with the University Open Days and advertised within the petroleum and mass spectrometry scientific communities (both academic and industrial) to maximize the impact which will have the result of increasing attendance and informing/attracting more and better PhD students. This publicity effort will be maximized by working with the Warwick University Communications Office.

We will make all developments (including circuit diagrams, code, CAD drawings, etc) available on our group website for access by anyone. We have a particularly good track record on this as evidenced by the 20+ groups who now have built copies of our RF oscillator design, the dozens of research groups who have downloaded and used BUDA (our open-source FTICR data analysis software), and two research groups which have copied our thin gate valve design.

As the project develops, we will ensure that commercial end users are aware of the developments both via our network of existing and past collaborators (Bruker, Exxon-Mobil, petroleum companies, etc) and through new external contacts that we are making through the Warwick Centre for Analytical Science (WCAS), which serves as a hub for analytical instrumentation development and application. This activity will ramp up as the project develops. To make certain that the economic impact of the developments made within this research are realized, commercializable developments that are derived from this research will be patented or otherwise IP protected and the Warwick academic staff and our industrial partners will engage with Warwick Ventures, Research Support Services, the Warwick Development Office, and Advantage West Midlands to ensure that these results are disseminated to and exploited by the business community.

Particularly, Drs. Baykut and Van Orden of Bruker, as project partners, will be closely involved in the research so that any developments can be directly implemented onto their instruments.