High Throughput Lipidomics Analysis by MALDI/Ion Mobility Mass Spectrometry

描述（由申请人提供）：代谢组学的研究领域包括对细胞内酶促反应产物和底物的广泛生物分子的研究。细胞代谢物的主要亚类之一是脂质，脂质本身是一种不同类型的分子，包括脂肪酸、类固醇、甘油脂、甘油磷脂、鞘脂和异戊烯醇。在过去的十年中，脂质的测量有了显著的改善，这主要是由于质谱和各种辅助技术的进步。脂质构成感兴趣的代谢组学领域内的一类重要化合物，因为这些分子不仅在通过形成脂质双层分离细胞内的区室中，而且通过这些分子可以作为信号分子的前体、过氧化化学的靶、DNA表达的调节剂、甚至是被蛋白质识别的实体，这些蛋白质驱动正常细胞生物学中的关键过程。在每类脂质中，存在称为分子种类的密切相关的分子，这增加了脂质混合物的复杂性和分析的挑战性。基于我们在使用MALDI成像质谱法方面取得的最新进展，我们建议将有可能显着增加脂质分析的通量，以便有可能在相当常规的基础上每天从数百个样品中获得重要的代谢组学信息。此外，我们建议，这个平台可以调整，以增加特定类别的脂质的灵敏度，并用于高通量lipidomic分析，以解决有关脂质合成和代谢中涉及的酶途径的具体问题。在某种程度上，这种方法将利用鸟枪脂质组学领域取得的进展，结合MALDI质谱的高通量能力，产生定性和定量信息。初步数据显示，即使是未经处理的血清和血浆也可以产生关于该液体中存在的脂质的复杂混合物的信息，而无需除稀释之外的任何进一步样品处理。该数据可以在不到一分钟的时间内以一种可以容易地集成到工具中的形式获得，以自动识别脂质以及定量存在的脂质的丰度。为了进行这些研究，将使用先进的质谱技术，包括离子迁移率技术，将脂质离子与MALDI电离过程中产生的其他化合物和杂质分离。即使没有离子迁移率，许多实验室也可以实施待开发的先进质谱技术;然而，随着离子迁移率变得越来越广泛，它提供了目前其他质谱系统所不具备的独特能力。

DESCRIPTION (provided by applicant): The area of research termed metabolomics encompasses the study of a wide range of biomolecules that are products and substrates of enzymatic reactions within cells. One of the major subclasses of cellular metabolites are lipids, which they themselves are a diverse class of molecules that range from fatty acids, steroids, glycerolipids, glycerophospholipids, sphingolipids, and prenols. The measurement of lipids has significantly improved over the past decade, largely due to advances in mass spectrometry and various ancillary techniques. Lipids constitute an important class of compounds within the metabolomic sphere of interest because of the central role these molecules play, not only in separating compartments within cells through formation of lipid bilayers, but also through specific roles these molecules can play as precursor of signally molecules, targets for peroxidation chemistry, regulators of DNA expression, and even entities that are recognized by proteins that drive critical processes in normal cell biology. Within each class of lipids, there ae closely related molecules termed molecular species which increase the complexity of the lipid mixture and challenging analysis. Based upon recent advances we have made in using MALDI imaging mass spectrometry we propose that it will be possible to significantly increase the throughput of lipid analysis so that it will be possible to obtain important metabolomic information from several hundreds of samples per day on a fairly routine basis. Furthermore, we propose that this platform can be tuned to increase sensitivity for specific classes of lipids and used for high throughput lipidomic analysis to address specific questions concerning enzymatic pathways involved in lipid synthesis and metabolism. In part this approach will take advantage of the advances that have been made in the area of shotgun lipidomics coupled to the high throughput capability of MALDI mass spectrometry to yield both qualitative and quantitative information. Preliminary data shows that even untreated serum and plasma can yield information about the complex mixture of lipids present in this fluid without any further sample treatment except dilution. This data can be obtained within less than a minute in a form that can readily integrate into tools to automatically identify lipids as well as quantitate the abundance o lipids present. In order to carry out these studies, advanced mass spectrometric techniques will be used, including ion mobility technology to separate lipid ions from other compounds and impurities generated during the MALDI ionization process. The advanced mass spectrometric techniques to be developed can be implemented in many laboratories even without the ion mobility; however, ion mobility, as it becomes more widely available, provides a unique level of capability not currently present with other mass spectrometric system.