Generation of antibodies to monitor Notch O-fucosylation in vivo

Project Summary Notch is a cell surface protein that has diverse roles in human biology at all stages of life Notch 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 Notch has significant effects on Notch function which in turn has been linked to both developmental and adult pathologies including cancer To date the study of O fucose structures on Notch 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 Notch 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 Notch 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 Notch If successful these O fucose Notch specific antibodies will facilitate research on Notch in many laboratories and possibly could lead to new diagnostic and therapeutic tools Project Narrative Modification of the human Notch protein by a sugar structure termed O fucose is now under scrutiny for its role in cell biology and human diseases such as cancer There are currently no protein specific reagents to study O fucose We propose to develop site specific O fucose antibodies to the human Notch protein thereby providing valuable tools for the study of O fucosylation and Notch biology and their roles in human disease