The development of methods to determine cell surface glycosylation with the rapid

描述(由申请人提供)：Glycomics正在迅速崛起，成为发现生物标记物的新范例。众所周知，各种疾病，如感染和癌症，都涉及糖基化的变化。细胞表面的葡聚糖对于理解几乎所有细胞表面的相互作用都很重要。它们是药物的关键靶点，可能会产生细胞特异性疗法。此外, 它们也会脱落，并能给出与疾病相关的糖基化变化的迹象。细胞表面糖基化的研究仍处于初级阶段。大多数研究使用了荧光标记的凝集素，提供的结构细节很少。在这项提案中，我们将开发研究表面多聚糖的技术，使用专门释放它们的方法，并以高灵敏度检查它们。在这个过程中，我们将开发全面的方法来确定多糖结构和微观不均质性。为了实现这些任务，我们将开发一种用于糖组分分析的高通量方法，该方法将快速识别多糖结构。考虑到多糖结构的复杂性和异质性，这一目标似乎是不可能的。然而，我们将开发一种以构建的数据库为核心的方法，该数据库将作为模板，描述符包括液相色谱保留时间、准确质量和串联质谱学，以鉴定单个多糖(或低聚糖)结构。这个数据库的建立将形成一个综合数据库的核心，使低聚糖的常规分析成为可能。快速鉴定方法的发展将极大地促进糖生物学研究。此外，通过分析结构异构体，用液相色谱分析多糖将为发现生物标记物提供一种新的方法，从而增加化合物池的丰富度。

DESCRIPTION (provided by applicant): Glycomics is rapidly emerging as a new paradigm for biomarker discovery. Diseases as diverse as infection and cancer are known to involve changes in glycosylation. Glycans on cell surfaces are important for understanding nearly all cell surface interactions. They are key targets for drugs and may yield cell-specific therapeutics. In addition, they are also shed and can give indications of the changes in glycosylation associated with the disease. The study of glycosylation of cell surfaces is still in its infancy. The majority of the research has employed fluorescently labeled lectins providing few structural details. In this proposal, we will develop techniques to study surface glycans by using methods that release them specifically and examine them with high sensitivity. In the process, we will develop comprehensive methods to determine glycan structures and micoheterogeneity. To achieve these tasks, we will develop a high throughput method for glycomics analysis that will rapidly identify glycan structures. This goal would have seemed impossible given the complexity and the heterogeneity of glycan structures. However, we will develop a method with a constructed database at its core that will serve as template with descriptors including liquid chromatography retention time, accurate mass, and tandem mass spectrometry to identify individual glycan (or oligosaccharide) structures. The creation of this database will form the kernel of a comprehensive database will allow routine analysis of oligosaccharide possible. The development of methods for the rapid identification will significantly advance glycobiology research. Additionally, the analysis of glycans by liquid chromatography will provide a new method for biomarker discovery by allowing the analysis of structural isomers thereby increasing the richness of the compound pool.