On-chip Glycan Analysis of Clinical Specimens

项目总结 最近的研究揭示了多糖对生物学的出人意料的贡献方式，以及新的策略 使用涉及多糖的方法与疾病作斗争。要充分利用这些机遇 首先，我们需要能够使非糖生物学专家的生物学家和临床医生 开展这一领域的研究。目前的方法需要高度的专业知识或专门的 设备。这些方法通常也不提供与疾病为导向的研究相关的信息，如 如临床标本中特定多糖的位置、性质和精确水平。这个项目的目标是 开发一种方法来帮助满足这一需求。这种方法，我们称之为芯片上葡聚糖修饰和 探测(片上GMAP)，非专家可以访问，样本量小且有效 在许多样品上提供精确的测量。我们之前演示了微尺度捕获和 基于亲和力的生物样品中多糖和糖蛋白的探测，以及算法和软件 用来解释数据。在这里，我们将添加捕获的葡聚糖的酶促变化，然后 凝集素/抗体探测改变的多糖和数据的自动解释。这些协议是 简单；试剂可以以试剂盒的形式提供；由于规模小，成本可以保持在较低水平； 而且，通过软件可以很容易地理解解释。这些信息将是 补充了传统的使用质谱学的糖链分析 层析法。质谱学和层析提供了组合物和候选结构 样本数量有限，新方法将以定量的方式提供许多 样本。因此，芯片上的GMAP可能成为实用和全面的葡聚糖的重要组成部分 分析策略，特别是在疾病研究方面。此外，它还可以扩大葡聚糖分析的用途 为不是糖生物学专家的研究人员提供工具，潜在地促进新的发现和 申请。

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.