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

SBIR 主题 294 第 1 阶段 - 糖基化的发展 - 特定研究

• 批准号: 8354002
• 负责人: LORI YANG
• 金额: $ 14.89万
• 依托单位: GLYCOSENSORS AND DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-15 至 2012-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8354002
• 关键词: Affinity; Antibodies; Antigen Targeting; Antigens; Binding; Biosensor; Biotin; Chromatography; Computing Methodologies; Detection; Development; Disease; Enzymes; Equilibrium; Escherichia coli; Exclusion; Glycopeptides; Glycoproteins; Homologous Gene; Human; Ion Exchange; Libraries; Malignant Neoplasms; Measures; Methods; Monosaccharides; Mutagenesis; Peptides; Phage Display; Phase; Polysaccharides; Preparation; Probability; Production; Proteins; Reagent; Regulation; Reporting; Research; Screening procedure; Site; Site-Directed Mutagenesis; Small Business Innovation Research Grant; Specificity; Surface Plasmon Resonance; System; Techniques; Testing; Tissues; Toxic effect; Variant; aptamer; base; c-myc Genes; cost; design; fast protein liquid chromatography; glycosylation; in vivo; innovation; instrument; protein aminoacid sequence; success; tool; vector

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.

该提案的长期目标是证明一种新的高亲和力高亲和力蛋白（称为Lectenz¿具体目的是创建一种试剂，该试剂可用于检测聚糖F3-0-GLCNAC本身，并在其疾病或癌症特异性肽序列（Glycosylation位点）中检测到能够检测F3-0-GICNAC的这种试剂的变化。迅速确认蛋白质和组织中F3-0-GLCNAC的存在的能力而无需转向更精致的技术，将为描绘差异糖基化的蛋白质提供强大的工具，并最终在F3-0-GICICNAC调控和相关疾病状态（例如癌症）之间建立相关性。使用点诱变，显示库筛选和最前沿的计算方法的组合，人酶O-GICNACASE（HOGA）将因其自然底物F3-0-GICNAC转换为高特异性生物传感器。 Lectenz®比抗体或适体具有多个潜在的优势，包括针对靶抗原的预定义特异性，以单价形式的易于制备，以及（对于人类同源物）体内毒性的可能性较低。计算方法的创新使用导致设计和筛选的效率提高 显示库，降低成本并提高成功率。