Making glycoproteomics via mass spectrometry more accessible to the greater scientific community

让广大科学界更容易利用质谱法进行糖蛋白组学

基本信息

批准号：
9893341
负责人：
Carolyn Bertozzi
金额：
$ 12.52万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9893341
关键词：
Adopted Area Benchmarking Biological Biological Assay Biological Process Cancer cell line Cell Culture Techniques Cells Chemistry Chromatography Code Collection Communities Complex Complex Mixtures Computer software Core Facility Country Data Data Files Databases Deposition Detection Disease Equipment Fractionation Glycobiology Glycopeptides Glycoproteins Goals Human Investigation Isotopes Knowledge Label Laboratories Libraries Malignant neoplasm of ovary Mass Spectrum Analysis Medical Metabolic Methodology Methods Online Systems Organic Synthesis Pattern Peptides Polysaccharides Protein Glycosylation Proteins Proteomics Protocols documentation Publishing Quality Control Reagent Research Research Personnel Resolution Sampling Services Shapes Site Standardization Structure System Techniques Technology Testing Time Vendor Work analytical tool anticancer research biological systems cancer diagnosis commercialization data warehouse glycoproteomics glycosylation improved instrument instrumentation interest mass spectrometer open source professor software development sugar tool web site

项目摘要

7. PROJECT SUMMARY/ABSTRACT We very recently published a method that, for the first time, allows glycan structures and sites of attachment to be discerned from complex biological samples for both N- and O-glycans alike. A major breakthrough is that this method does not require truncation of the glycans, allowing the rich glycomic information to remain intact during analysis. In addition, this method, termed Isotope-Targeted Glycoproteomics (IsoTaG) enriches glycopeptides and allows unprecedented detection of low abundance glycoproteins. IsoTaG also mitigates the need for extensive fractionation, mass spectrometer analysis time, and computation time. Also, we have since been able to transfer the IsoTag method to interested labs that have no prior glycobiology expertise, who were able to successfully perform glycoproteomic analysis on their samples of interest. With these barriers to large scale implementation of glycoproteomics substantially reduced, we propose herein to make IsoTaG accessible to the broader scientific community as a standard service offered by mass spectrometry (MS) core facilities. To accomplish this, we will improve the IsoTaG reagent and develop a large scale synthesis to enable distribution to the community. We will also standardize various aspects of the chromatography used to separate glycopeptides, including making standards that can be used to calibrate retention times. Also, we will develop protocols for use of these standards to tune instrument parameters for optimal fragmentation of the glycopeptides. Finally, we will provide the reagents and protocols to several mass spectrometry core facilities to trial use of the IsoTag system on their instruments. Ensuing dialog with the facilities will help us improve the method with the goal of it being adoptable by any facility that has high resolution MS instrumentation.

7。项目摘要/摘要我们最近发表了一种方法，该方法首次允许聚糖结构和附着的位置对于N-和O-Glycans，都可以从复杂的生物样品中辨别。一个主要的突破是此方法不需要截断聚糖，允许丰富的糖果信息保持完整在分析过程中。另外，该方法称为同位素靶向糖蛋白质组学（ISOTAG）富集糖肽并允许对低丰度糖蛋白的前所未有的检测。同构也会减轻需要大量分馏，质谱仪分析时间和计算时间。而且，我们从那以后能够将Isotag方法转移到没有先前没有糖生物学专业知识的有兴趣的实验室能够成功地对其感兴趣的样本进行糖蛋白质组学分析。这些障碍糖蛋白质组学的规模实施大大降低作为质谱（MS）核心设施提供的标准服务，更广泛的科学界。为此，我们将改进同塔试剂并开发大规模合成以实现向社区分发。我们还将标准化色谱的各个方面单独的糖肽，包括制定可用于校准保留时间的标准。另外，我们会的开发用于使用这些标准来调整仪器参数的协议糖肽。最后，我们将为几个质谱核心设施提供试剂和协议试用在其仪器上使用同位物质系统。随后与设施的对话将有助于我们改善任何具有高分辨率MS仪器的设施都可以采用的方法。