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

该提案的长期目标是证明一类称为Lectenz的新型高特异性高亲和力蛋白质作为用于识别疾病或癌症相关糖基化位点的研究试剂的实用性。具体目的是创建一种用于检测聚糖f3-0-GlcNAc本身的泛特异性试剂，以及能够在其疾病或癌症特异性肽序列（糖基化位点）的背景下检测f3-0-GlcNAc的该试剂的变体。快速确认蛋白质和组织中f3-0-GlcNAc的存在而不需要转向更精细的技术的能力将提供描绘差异糖基化蛋白质并最终建立f3-0-GlcNAc调节与相关疾病状态（例如癌症）之间的相关性的有力工具。使用点突变、展示文库筛选和前沿计算方法的组合，人酶O-GlcNAc酶（hOGA）将被转化为其天然底物f3-0-GlcNAc的高特异性生物传感器。莱滕茨与抗体或适体相比具有几个潜在的优点，包括预定的靶抗原特异性、易于以单价形式制备和（对于人同源物）体内毒性的可能性低。计算方法的创新使用导致在设计和筛选的效率增益 显示库，降低成本，提高成功率。

The long term objectives of this proposal are to demonstrate the utility of a new class of high-specificity high-affinity proteins called Lectenz¿ as research reagents for use in recognition of disease- or cancer-related glycosylation sites. The specific aims are to create a reagent that is pan-specific for the detection of the glycan f3-0-GlcNAc itself, as well as variations of this reagent that are able to detect f3-0-GIcNAc in the context of its disease- or cancer-specific peptide sequence (glycosylation site). The ability to rapidly confirm the presence of f3-0-GlcNAc in proteins and tissues without the need to turn to more-elaborate techniques would provide a powerful tool to delineate differentially glycosylated proteins and eventually establish correlations between f3-0-GIcNAc regulation and associated disease states, such as cancer. Using a combination of point mutagenesis, display library screening, and forefront computational methods, the human enzyme O-GIcNAcase (hOGA) will be converted into a high-specificity biosensor for its natural substrate f3-0-GIcNAc. Lectenz¿ have several potential advantages over antibodies or aptamers, including predefined specificity for the target antigen, ease of preparation in a monovalent form, and (for human homologs) a low probability of in vivo toxicity. The innovative use of computational methods results in efficiency gains in the design and screening of the display library, lowering costs and increasing success rates.