Generation of antibodies to monitor Notch O-fucosylation in vivo

生成用于监测体内 Notch O-岩藻糖基化的抗体

• 批准号: 9200115
• 负责人: RON ORLANDO
• 金额: $ 24.23万
• 依托单位: GLYCOSCIENTIFIC, LLC
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-26 至 2018-09-25
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9200115
• 关键词: Adult; Amino Acid Sequence; Antibodies; Antibody Formation; Antibody Specificity; Binding; Biological; Biological Markers; Biology; Biopsy; Cell Surface Proteins; Cells; Cellular biology; Clinical; Communities; Cultured Cells; Defect; Detection; Development; Disaccharides; Disease; Disease Marker; Epidermal Growth Factor; Epitopes; Extracellular Domain; Family; Fucose; Fucosyltransferase 1; Generations; Human; Human Biology; Immunization; Individual; Laboratories; Lead; Life; Link; Malignant Neoplasms; Mass Spectrum Analysis; Methodology; Methods; Modification; Monitor; Monoclonal Antibodies; Monosaccharides; N-Acetylglucosaminyltransferases; Pathology; Pattern; Peptides; Phase; Polysaccharides; Post-Translational Protein Processing; Production; Proteins; Reaction; Reagent; Research; Research Personnel; Role; Sampling; Site; Specificity; Specimen; Staging; Structure; Techniques; Time; Tissues; cost effective; glycosylation; human disease; human tissue; in vivo; instrumentation; interest; malignant breast neoplasm; meetings; notch protein; novel; novel diagnostics; novel therapeutics; polyclonal antibody; sugar; tool

Project Summary Notch1 is a cell surface protein that has diverse roles in human biology at all stages of life. Notch1 is modified by a number of sugar structures, including a number that are termed O-fucose. Alteration of the pattern of O- fucose structures found on Notch1 has significant effects on Notch1 function, which in turn has been linked to both developmental and adult pathologies, including cancer. To date the study of O-fucose structures on Notch1 has been severely hindered by a lack of research reagents that can specifically detect and monitor this structure. Currently, mass spectrometry is the method of choice for the analysis of O-fucose glycans on Notch1. Although an extremely powerful technique, it requires specialized instrumentation and expertise not available in most laboratories. Development of sensitive and specific antibodies for O-fucose glycans on Notch1 will provide many investigators with the tools to analyze O-fucose glycans in vivo, including clinical specimens. We will utilize a newly-developed methodology that enables reliable production of antibodies that can recognize distinct protein epitopes in the context of specific glycan modifications to produce antibodies to O- fucose on Notch1. If successful these O-fucose-Notch1-specific antibodies will facilitate research on Notch1 in many laboratories and possibly could lead to new diagnostic and therapeutic tools.

项目摘要 Notch1是一种细胞表面蛋白，在生命的各个阶段都在人类生物学中具有不同的作用。 Notch1已修改 通过许多糖结构，包括一个称为O型凝血的数字。 O-模式的改变 Notch1上发现的岩藻糖结构对Notch1功能具有重大影响，而Notch1功能又与 发育和成人病理，包括癌症。迄今为止的O谷糖结构的研究 缺乏可以专门检测和监测的研究试剂的缺乏研究试剂严重阻碍了Notch1 结构。 当前，质谱是在Notch1上分析O羟基聚糖的首选方法。虽然 一种非常强大的技术，它需要大多数专业的仪器和专业知识 实验室。在Notch1上开发O羟基聚糖的敏感和特异性抗体将提供 许多研究人员使用的工具可以分析体内的O-糖粉聚糖，包括临床标本。 我们将利用一种新开发的方法，该方法可实现可靠的抗体生产 在特定的聚糖修饰的背景下，识别不同的蛋白质表位，以产生与O-的抗体 Notch1上的Fucose。如果成功这些O-凝血核-Notch1特异性抗体将有助于对Notch1进行研究 许多实验室，可能会导致新的诊断和治疗工具。