SBIR TOPIC 294 PHASE 1 - DEVELOPMENT OF GLYCOSYLATION - SPECIFIC RESEARCH

碳水化合物修饰O-GlcNAc几乎与蛋白质中的蛋白质磷酸化一样丰富，与其竞争，并具有广泛的串扰以调节信号传导、转录以及癌基因和肿瘤抑制因子的功能。然而，蛋白质O-GlcNAc化的分析目前需要困难且耗时的方法，例如复杂的质谱法。有必要开发新的试剂和更简单的方法来研究癌症生物学中蛋白质的碳水化合物修饰（O-连接和N-连接）的功能。 即使在有能力制造糖基化特异性抗体的实验室中，在对该领域产生影响的规模上做这件事也太耗时和昂贵了。另一方面，DNA适体易于合成，并且比抗体便宜至少10倍。 该项目利用一种新的工具，即DNA适体的平行选择，同时快速鉴定数十甚至数百种糖蛋白或修饰多肽靶点的高亲和力配体。在这个NIH I期SBIR项目中，我们将展示我们的工具用于选择与癌症生物学有关的许多重要糖蛋白的适体的实用性。我们还将开发针对特定糖肽的高亲和力试剂，这些试剂可能在癌症糖生物学的理解中发现巨大的效用。

The carbohydrate modification O-GlcNAc is nearly as abundant as protein phosphorylation in proteins, competes with it, and has extensive crosstalk to regulate signaling, transcription, and the functions of oncogenes and tumor suppressors. However, analysis of protein O-GlcNAcylation currently requires difficult and time-consuming methods such as sophisticated mass spectrometry. There is a need to develop new reagents and simpler methods to study the function of carbohydrate modifications (O-linked and N-linked) of proteins in cancer biology.¿ Even in labs with the ability to make glycosylation-specific antibodies, it is simply too time-consuming and expensive to do that on a scale which would have an impact on the field. DNA aptamers, on the other hand, are readily synthesized and at least 10 times less expensive than antibodies. This project utilizes a new tool, ¿parallel selection of DNA aptamers¿ to rapidly identify high-affinity ligands to tens or even hundreds of glycoprotein or modified polypeptide targets simultaneously. In this NIH Phase I SBIR project, we will demonstrate the utility of our tool for selecting aptamers to numerous important glycoproteins implicated in cancer biology. We will also develop high affinity reagents to specific glycopeptides which are likely to find enormous utility in the understanding of cancer glycobiology.