Generation of antibodies to monitor Notch O-fucosylation in vivo

项目摘要 NOTCH1是一种细胞表面蛋白，在人类生命的所有阶段都有不同的生物学作用。NOTCH1已修改 由许多糖结构组成，包括一些被称为O-岩藻糖的糖。O-句式的变化 Notch1上发现的岩藻糖结构对Notch1功能有显著影响，而Notch1功能又与 包括癌症在内的发育和成人病理。O-岩藻糖结构的研究进展 NOTCH1由于缺乏专门检测和监测这一现象的研究试剂而受到严重阻碍 结构。 目前，质谱法是分析Notch1上O-岩藻糖多糖的首选方法。虽然 作为一种非常强大的技术，它需要专门的仪器和专业知识，这是大多数 实验室。针对Notch1上O-岩藻糖聚糖的敏感和特异抗体的开发将提供 许多研究人员利用该工具分析了O-岩藻糖在体内的多糖，包括临床标本。 我们将利用一种新开发的方法，使可靠的抗体生产能够 在特定的糖链修饰背景下识别不同的蛋白质表位以产生O-抗体 Notch1上的岩藻糖。如果成功，这些O-岩藻糖-Notch1特异性抗体将促进对Notch1的研究 许多实验室，并可能导致新的诊断和治疗工具。

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.