On-chip Glycan Analysis of Clinical Specimens

项目摘要 最近的研究发现了聚糖对生物学的意想不到的贡献，以及新的策略 用聚糖来对抗疾病。充分利用这些机会， 开放，我们需要的方法，将使生物学家和临床医生谁不是专家在糖生物学， 在这个领域进行研究。目前的方法需要高度的专业知识或专门知识， 设备.这些方法通常也不提供与疾病导向研究相关的信息，例如 如临床标本中特定聚糖的位置、性质和精确水平。该项目的目标是 开发一种方法来帮助满足这种需求。我们称之为芯片上聚糖修饰的方法， 探测（片上GMAP），将可供非专业人员使用，可用于低样本量， 在许多样品上提供精确的测量。我们之前展示了微尺度捕获和 生物样品中聚糖和糖蛋白的基于亲和力的探测，以及算法和软件 来解释数据。在这里，我们将添加捕获聚糖的酶促改变，然后 凝集素/抗体探测改变的聚糖和数据的自动解释。协议是 简单明了;试剂可以试剂盒形式提供;由于小型化规模，成本可以保持较低; 并且可以通过软件容易地理解解释。这些信息将被 与使用质谱法从常规聚糖分析得到的结果互补， 层析质谱法和色谱法提供了一种新的生物样品的组成和候选结构。 有限数量的样本，新方法将提供基序信息，在定量的方式超过许多 样品因此，芯片上GMAP可能是实用和全面的聚糖的重要组成部分 分析策略，特别是疾病研究。此外，它可以拓宽聚糖分析的用途 为非糖生物学专家的研究人员提供工具，可能促进新的发现， 应用.

PROJECT SUMMARY Recent research is uncovering unexpected ways that glycans contribute to biology, as well as new strategies for combatting diseases using approaches involving glycans. To make full use of the opportunities that are opening, we need methods that will enable biologists and clinicians who are not experts in glycobiology to conduct research in this field. The current methods require a high degree of expertise or specialized equipment. The methods also typically do not provide information relevant to disease-oriented research, such as the location, nature, and precise levels of specific glycans in clinical specimens. The goal of this project is to develop a method that will help meet this need. The method, which we call On-Chip Glycan Modification and Probing (On-Chip GMAP), will be accessible to non-specialists, usable with low sample volumes, and effective in providing precise measurements over many samples. We previously demonstrated micro-scale capture and affinity-based probing of glycans and glycoproteins from biological samples, as well as algorithms and software for interpreting the data. Here we will add enzymatic alterations of the captured glycans, followed by lectin/antibody probing of the altered glycans and automated interpretation of the data. The protocols are straightforward; reagents can be supplied in kit form; costs can be kept low because of the miniaturized scale; and the interpretation can be made readily understandable through the software. The information will be complementary to that derived from conventional glycan analyses using mass spectrometry and chromatography. Mass spectrometry and chromatography provide compositions and candidate structures for a limited number of samples, and the new method will provide motif information in a quantitative way over many samples. Therefore, On-Chip GMAP could be an important component of practical and comprehensive glycan analysis strategies, particularly for disease research. In addition, it could broaden the use of glycan analysis tools to researchers who are not experts in glycobiology, potentially facilitating new discoveries and applications.