On-chip Glycan Analysis of Clinical Specimens

临床样本的片上聚糖分析

• 批准号: 9333187
• 负责人: Brian B. Haab
• 金额: $ 30.46万
• 依托单位: VAN ANDEL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-16 至 2018-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9333187
• 关键词: Address; Affinity; Algorithmic Software; Algorithms; Antibodies; Area; Binding; Binding Proteins; Biochemistry; Biological; Biological Assay; Biology; Cell surface; Clinical; Complex; Computer software; Consumption; Data; Data Analyses; Disease; Enzyme-Linked Immunosorbent Assay; Enzymes; Equipment; Evaluation; Future; Glycobiology; Glycoproteins; Goals; Knowledge; Lectin; Location; Mass Chromatography; Mass Spectrum Analysis; Measurement; Methods; Modification; Nature; Polysaccharides; Population; Proteins; Protocols documentation; Reagent; Reproducibility of Results; Research; Research Personnel; Sampling; Series; Software Tools; Specificity; Specimen; Structure; Technology; Vision; Work; base; combat; computerized tools; cost; experimental study; glycosylation; miniaturize; software development; technology development; tool

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.

项目概要 最近的研究揭示了聚糖对生物学的贡献的意想不到的方式，以及新的策略 使用涉及聚糖的方法来对抗疾病。充分利用现有的机会 首先，我们需要一些方法，使非糖生物学专家的生物学家和临床医生能够 进行该领域的研究。当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导向研究相关的信息，例如 作为临床样本中特定聚糖的位置、性质和精确水平。该项目的目标是 开发一种有助于满足这一需求的方法。我们将这种方法称为片上聚糖修饰（On-Chip Glycan Modification） 探测（片上 GMAP）可供非专业人士使用，可用于低样本量且有效 提供对许多样品的精确测量。我们之前演示了微尺度捕获和 基于亲和力的生物样品中聚糖和糖蛋白的探测以及算法和软件 用于解释数据。在这里，我们将添加捕获的聚糖的酶促改变，然后 凝集素/抗体探测改变的聚糖并自动解释数据。协议是 直截了当;试剂可以以试剂盒形式提供；由于规模小型化，成本可以保持在较低水平； 并且通过软件可以使解释变得容易理解。该信息将是 与使用质谱法进行的传统聚糖分析的补充 色谱法。质谱和色谱法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的主题信息 样品。因此，On-Chip GMAP 可能成为实用和综合聚糖的重要组成部分 分析策略，特别是疾病研究。此外，它还可以扩大聚糖分析的用途 为非糖生物学专家的研究人员提供工具，有可能促进新的发现和 应用程序。