DEVELOPMENT & UTILIZATION OF A MUCIN-GLYCOPEPTIDE MICROARRAY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 由于粘蛋白结构域是许多细胞表面分子的组成部分，而对这些结构的识别在细胞发育和疾病中非常重要，因此我们有兴趣开发有效的方法来识别它们的伙伴和相互作用。粘蛋白糖蛋白的下层支架在展示悬挂的糖蛋白表位时的结构特征是重要的。因此，为了定量地描述它们之间的相互作用，需要相关的糖共轭结构，而不是孤立的多糖。我们的目标是通过开发微阵列平台来满足这一需求，该平台将允许为合作伙伴进行快速筛选。在此类研究中，此类结构的可用性一直是一个限制。我们采用了一种基于糖肽化学合成的方法，提供了定义良好的结构，可以进一步从生物化学的角度进行阐述。在开发一种强大的阵列时，正在开发和评估用于将糖肽锚定到载玻片上的新型连接物和连接方法。最初选择的分子将显示截短的糖链，就像在肿瘤相关糖蛋白中发现的那样。 将使用针对印刷结构的单克隆和多克隆抗体来评估阵列上的分子展示的细节。由我们的化学方法产生的糖结合物上的糖基化的系统变化将允许定义相互作用的选择性。这将扩展到对患者血清样本的筛选，以研究其在检测抗体癌症生物标志物方面的应用。该项目汇集了佐治亚大学CCRC在化学合成和结构分析方面的专业知识，以及埃默里大学糖学中心与多糖相关的阵列专业知识。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Because mucin domains are components of a number of cell surface molecules, and recognition of these structures are important in cellular development and disease, we are interested in developing efficient methods for identifying their partners and interactions. Structural characteristics of the underlying scaffold of mucin glycoproteins in the display of pendant glycan epitopes are important. Therefore, to quantitatively characterize their interactions, relevant glycoconjugate constructs are needed rather than isolated glycans. Our goal is to address the requirement through development of a microarray platform that will allow rapid screening for partners. Availability of such constructs has been a limitation in such studies. We have adopted an approach based on glycopeptide chemical synthesis, providing well-defined structures, which can be further elaborated biochemically. In developing a robust array, novel linkers and ligation approaches are being developed and evaluated for anchoring glycopeptides to the slide. The initial selection of molecules will display truncated glycans as found in tumor associated glycoproteins. Details of molecular display on the array will be evaluated using monoclonal and polyclonal antibodies to printed structures. Systematic variation of the glycosylation on the glycoconjugates resulting from our chemical approach will allow definition of the selectivity of the interactions. This will be extended to screening of patient serum samples to investigate applications to detection of antibody cancer biomarkers. The project brings together the expertise in chemical synthesis and structural analysis at the CCRC at the University of Georgia, and glycan related array expertise at the Glycomics Center at Emory University.