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

这项研究的目的是开发更好的仪器，这将有助于各种各样的化学和生化问题，如了解疾病的基本机制和石油副产品通过地下水的迁移。例如，1型糖尿病是一种疾病，某些事件触发身体的免疫系统杀死胰腺中产生胰岛素的胰岛细胞。这种病毒产生的触发这种免疫反应的确切分子变化也不清楚，但它通常刺激产生针对胰岛素本身、胰岛素前体蛋白或胰岛细胞表面蛋白（如GAD65）的抗体。为了确切地了解问题是什么，从而能够开发更有效的治疗方法，并有望治愈，弄清楚这种确切变化的本质是很重要的。不幸的是，这是一个典型的“大海捞针”问题。大多数组织中有成千上万种蛋白质，尽管医学界/科学界对所涉及的蛋白质和对它们进行的修饰有许多很好的猜测，但样本仍然非常复杂。分离这些混合物的大部分成分是可能的，但即使候选分子被分离出来，也有必要对它们进行完整的测序，而不破坏修饰，以便确切地找出变化。质谱法可以做到这一点，但它们都有局限性。这项研究进展旨在开发一种新的、公正的方法，在复杂混合物中完全分割蛋白质和其他生物分子，而无需预先选择要关注的生物分子。通过同时生成混合物中所有成分的碎片质量信息，可以更快地生成更多数据，并且偏差很小，然后可以对重要信息进行数据挖掘。另一个例子涉及追踪炼油厂石油副产品对地下水（和鱼类）的潜在污染来源，以及测试生物修复方法。在此开发的相位控制方法将提高我们在许多样品中确定与油有关的化合物的存在和结构的能力，从土壤到地下水，再到生物修复藻类。所涉及的主要方法，称为二维傅立叶变换（2DFT）质谱法，是核磁共振相关光谱方法（如NOESY）的必然结果。2DFT方法实际上在20世纪80年代末就已经出现了，但在当时是不切实际的。这种不切实际的情况是当时使用的分散方法的副作用，但可以通过使用过去十年中已经得到甚至广泛使用的新方法来避免。此外，这种方法可以潜在地应用于各种离子，而不需要对控制电压、电流和激光功率进行太多的修改。如果这一前景取得成果，就可以设计和制造新的仪器，使大多数生物医学研究人员能够常规地进行这种分析。当然，这种方法并不局限于糖尿病和石油污染的研究，而是可以用于研究任何分子的任何修饰，无论是与心脏病还是炭疽疫苗有关。因此，它将在制药、化工、石油和生物技术行业的开发、质量控制、跟踪和故障排除方面有很大的用途。理论上，它可以应用于任何复杂的混合物，只要这种分子的破碎是有效的。一如既往，对于科学来说，细节决定成败，在这个项目中开发的方法将产生比以前更详细的复杂样本信息。

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.