Development of a Carbohydrate Microarray

碳水化合物-蛋白质相互作用对于许多生物学过程是必不可少的。 过程，如炎症、细菌和病毒粘附以及转移。因此，在本发明中， 人们对鉴定碳水化合物结合蛋白很感兴趣， 开发配体以调节它们的活性。碳水化合物-蛋白质相互作用分析 是由很多因素造成的首先，碳水化合物极难分离 或合成。因此，在许多情况下只能获得少量。第二、 用于研究结合的传统方法不是高通量的并且需要大量的 材料第三，碳水化合物和蛋白质之间的单价相互作用是众所周知的。 弱.为了获得高亲合力，碳水化合物结合蛋白含有多个结合位点 并形成多价复合物。因此，碳水化合物必须以多价形式存在， 方式和碳水化合物的间距和方向在 识别.碳水化合物微阵列，或聚糖阵列，包含许多不同的碳水化合物， 或聚糖，以空间限定的排列固定在固体支持物上。使用数组， 可以评价凝集素、抗体、细胞或病毒与糖链上所有碳水化合物的结合， 数组同时该阵列提供碳水化合物的多价呈现，并且 小型化形式仅需要皮克量的每种碳水化合物用于测定。我们有 开发了一种利用新糖蛋白的新型聚糖阵列形式。新糖蛋白是 多价缀合物，其中碳水化合物与载体蛋白共价连接， 如牛血清白蛋白。这些新糖蛋白，以及天然糖蛋白， 使用机器人微阵列仪打印到环氧涂层的显微镜载玻片上， 微阵列新糖蛋白可用作用于其它测定的多价试剂， ELISA，作为多价抑制剂，和作为多价免疫原。我们目前有一个 世界上最大最独特的聚糖阵列最近，我们专注于扩大和 改进碳水化合物微阵列。一个目标是增加聚糖的数量， 阵我们一直在开发用于制备糖肽的改进的合成方法。的 方法学将用于制备大的糖肽文库，以添加到 阵此外，我们正在优化将碳水化合物与蛋白质结合的方法， 产生新糖蛋白。

Carbohydrate-protein interactions are essential for a wide range of biological processes such as inflammation, bacterial and viral adhesion, and metastasis. As a result, there has been significant interest in identifying carbohydrate binding proteins and developing ligands to modulate their activity. Analysis of carbohydrate-protein interactions is complicated by a number of factors. First, carbohydrates are extremely difficult to isolate or synthesize. Therefore, only small amounts can be obtained in many cases. Second, traditional methods for studying binding are not high-throughput and require large amounts of material. Third, monovalent interactions between carbohydrates and proteins are notoriously weak. To achieve a high avidity, carbohydrate-binding proteins contain multiple binding sites and form multivalent complexes. As a result, carbohydrates must be presented in a multivalent fashion and the spacing and orientation of the carbohydrates play a critical role in recognition. Carbohydrate microarrays, or glycan arrays, contain many different carbohydrates, or glycans, immobilized on a solid support in a spatially-defined arrangement. Using an array, one can evaluate binding of a lectin, antibody, cell, or virus to all the carbohydrates on the array simultaneously. The array provides a multivalent presentation of carbohydrates, and the miniaturized format requires only picogram amounts of each carbohydrate for an assay. We have developed a novel glycan array format utilizing neoglycoproteins. Neoglycoproteins are multivalent conjugates wherein carbohydrates are covalently attached to a carrier protein, such as bovine serum albumin. These neoglycoproteins, as well as natural glycoproteins, are printed onto epoxide coated microscope slides using a robotic microarrayer to produce microarrays. Neoglycoproteins can be used as multivalent reagents for other assays such as ELISAs, as multivalent inhibitors, and as multivalent immunogens. We currently have one of the largest and most unique glycan arrays in the world. Recently, we have focused on expanding and improving the carbohydrate microarray. One objective is to increase the number of glycans on the array. We have been developing improved synthetic methods for preparing glycopeptides. The methodology will be used to prepare a large library of glycopeptides for addition to the array. in addition, we are optimizing methods to conjugate carbohydrates to proteins to produce neoglycoproteins.