Generation of antibodies to monitor Notch O-fucosylation in vivo
Generation of antibodies to monitor Notch O-fucosylation in vivo
批准号:
9200115
负责人:
RON ORLANDO
金额:
$24.23万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
2016
资助国家:
美国
项目状态:
已结题
起止时间:
2016-09-26 至 2018-09-25
关键词:
AdultAmino Acid SequenceAntibodiesAntibody FormationAntibody SpecificityBindingBiologicalBiological MarkersBiologyBiopsyCell Surface ProteinsCellsCellular biologyClinicalCommunitiesCultured CellsDefectDetectionDevelopmentDisaccharidesDiseaseDisease MarkerEpidermal Growth FactorEpitopesExtracellular DomainFamilyFucoseFucosyltransferase 1GenerationsHumanHuman BiologyImmunizationIndividualLaboratoriesLeadLifeLinkMalignant NeoplasmsMass Spectrum AnalysisMethodologyMethodsModificationMonitorMonoclonal AntibodiesMonosaccharidesN-AcetylglucosaminyltransferasesPathologyPatternPeptidesPhasePolysaccharidesPost-Translational Protein ProcessingProductionProteinsReactionReagentResearchResearch PersonnelRoleSamplingSiteSpecificitySpecimenStagingStructureTechniquesTimeTissuescost effectiveglycosylationhuman diseasehuman tissuein vivoinstrumentationinterestmalignant breast neoplasmmeetingsnotch proteinnovelnovel diagnosticsnovel therapeuticspolyclonal antibodysugartool
中文摘要
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。
英文摘要
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.
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