Generation of antibodies to monitor Notch O-fucosylation in vivo

项目摘要 Notch 1是一种细胞表面蛋白，在生命的各个阶段的人类生物学中具有不同的作用。Notch 1被修改 由许多糖结构组成，包括一些称为O-岩藻糖的结构。O-型的改变 在Notch 1上发现的岩藻糖结构对Notch 1功能有显著影响，这反过来又与Notch 1的功能有关。 包括癌症在内的发育和成人病理学。迄今为止，O-岩藻糖结构的研究 Notch 1由于缺乏能够特异性检测和监测Notch 1的研究试剂而受到严重阻碍。 结构 目前，质谱法是Notch 1上O-岩藻糖聚糖分析的首选方法。虽然 这是一种非常强大的技术，它需要专业的仪器和专业知识，而大多数国家都没有。 laboratories.开发针对Notch 1上的O-岩藻糖聚糖的敏感性和特异性抗体将提供 许多研究人员使用工具分析体内O-岩藻糖聚糖，包括临床标本。 我们将利用一种新开发的方法，能够可靠地生产抗体， 在特异性聚糖修饰的背景下识别不同的蛋白质表位，以产生针对O- Notch 1上的岩藻糖。如果成功的话，这些O-岩藻糖-Notch 1特异性抗体将促进对Notch 1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.