Novel Materials and Methods for Separations of Glycopeptides and Glycans
用于分离糖肽和聚糖的新材料和方法
基本信息
- 批准号:8899355
- 负责人:
- 金额:$ 21.91万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2011
- 资助国家:美国
- 起止时间:2011-05-01 至 2016-11-30
- 项目状态:已结题
- 来源:
- 关键词:Biological ProcessBiologyCarbohydratesCharacteristicsChemicalsChemistryCommunicable DiseasesComplexComplex MixturesDiseaseDisease MarkerExhibitsExperimental DesignsGlycopeptidesGlycoproteinsGoalsHigh Pressure Liquid ChromatographyHourInfluenzaKineticsKnowledgeLiquid ChromatographyMalignant NeoplasmsMarketingMass Spectrum AnalysisMeasuresMethodsModificationNatureOutcomePatientsPeptidesPerformancePhasePolysaccharidesPost-Translational Protein ProcessingProceduresPropertyProtein GlycosylationProteinsProteomicsResolutionSamplingScienceSeriesSilicon DioxideSiteSpeedStructureSurfaceSurface PropertiesTechnologyTestingThickTimeWorkanalytical toolcostdesignglycosylationimprovedliquid chromatography mass spectrometrymass spectrometermeetingsnanoparticlenew technologynoveloperationparticlepolypeptideprotein structurepublic health relevancereal world applicationresearch studysmall moleculesugartrend
项目摘要
DESCRIPTION (provided by applicant): Analysis of glycoprotein polypeptide sites of carbohydrate attachment and the structures of the glycans present on these proteins requires the combination of high resolution chromatographic separations and sophisticated mass spectrometry. Hydrophilic interaction liquid chromatography (HILIC) has great use in separations of glycopeptides and released glycans, exhibiting remarkable selectivity for the separations of glycoforms. The proposed effort is to markedly improve separations of glycopeptides and released glycans by HILIC, reducing the separation times from the current 2-3 hours per sample, to 30 minutes or less. The separations component of LC/MS analysis of glycoproteins can benefit from the design of more efficient stationary phase chromatographic materials, and from the use of conditions that allow direct and productive interfacing with mass spectrometers. We propose that recently developed superficially-porous silica microparticulate silica packing materials (Fused-Core(R) structures) can achieve these separation speeds, without loss of the required high resolution. The AMT Fused-Core reversed phase materials have previously been shown to exhibit superior kinetic properties and column efficiencies for separations of small molecules, and recently, wider pore size packings have shown a similar benefit for larger peptides (c. 3-5 kDa). The current proposal uses Fused-Core technology with unique surface modifications to develop high performance HILIC materials specifically for separations of glycopeptides and glycans. Tailored HILIC materials will be synthesized and tested for HILIC operation, as well as to establish the fundamental relationships between surface properties, particle characteristics, and utility for glycan and glycopeptides separations.
The new bonded phases utilize organosilane chemistry discovered in Phase I efforts that have been selected to withstand aggressive conditions of use, permitting stable and robust separations materials that can be used across a broad range of conditions. Preliminary LC/MS applications in glycoproteomic applications have been completed in Phase. A reasonable estimate is that an improvement of >2-fold for separations times are obtained. Phase II will expand on this effort, with the purpose of delivering further improved materials and methods to a broader range of needs in glycoproteomics and proteomics workflows. A significant aspect of the current proposal will assess the benefits and real-world applications of these new separations materials. The target of this work is to produce robust materials that permit simple integration with online mass spectrometry analysis, effective for resolution of the complex mixtures that are typical of current glycoprotein identification and structural characterization procedures.
描述(由申请方提供):糖蛋白多肽的碳水化合物连接位点和这些蛋白质上存在的聚糖结构的分析需要高分辨率色谱分离和复杂的质谱分析相结合。亲水相互作用液相色谱(HILIC)在糖肽和糖链的分离中有着广泛的应用,对糖型的分离具有显著的选择性。建议的努力是显着改善分离的糖肽和释放的聚糖HILIC,减少分离时间从目前的2-3小时,每个样品,到30分钟或更少。分离成分的LC/MS分析的糖蛋白可以受益于更有效的固定相色谱材料的设计,并从使用的条件,允许直接和生产性的接口与质谱仪。我们提出,最近开发的表面多孔二氧化硅微粒二氧化硅填充材料(Fused-Core(R)结构)可以实现这些分离速度,而不会损失所需的高分辨率。AMT融合核反相材料先前已经显示出对于小分子的分离表现出上级动力学性质和柱效率,并且最近,更宽孔径的填料已经显示出对于较大肽的类似益处(c. 3-5 kDa)。目前的提案使用具有独特表面改性的融合核技术来开发专门用于糖肽和聚糖分离的高性能HILIC材料。定制的HILIC材料将被合成并测试HILIC操作,以及建立表面性质,颗粒特性和聚糖和糖肽分离的实用性之间的基本关系。
新的键合相利用了在第一阶段工作中发现的有机硅烷化学,这些有机硅烷化学已经被选择为能够承受侵蚀性的使用条件,从而允许稳定和稳健的分离材料,这些分离材料可以在广泛的条件下使用。LC/MS在糖蛋白质组学应用中的初步应用已在第三阶段完成。合理的估计是,获得了>2倍的分离时间的改进。第二阶段将扩大这一努力,目的是提供进一步改进的材料和方法,以满足糖蛋白质组学和蛋白质组学工作流程中更广泛的需求。 当前提案的一个重要方面将评估这些新型分离材料的好处和实际应用。这项工作的目标是生产耐用的材料,允许与在线质谱分析简单集成,有效地解析当前糖蛋白鉴定和结构表征程序中典型的复杂混合物。
项目成果
期刊论文数量(2)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Optimized superficially porous particles for protein separations.
- DOI:10.1016/j.chroma.2013.09.054
- 发表时间:2013-11-08
- 期刊:
- 影响因子:0
- 作者:Schuster SA;Wagner BM;Boyes BE;Kirkland JJ
- 通讯作者:Kirkland JJ
Predicting the Retention Behavior of Specific O-Linked Glycopeptides.
- DOI:10.7171/jbt.17-2803-003
- 发表时间:2017-09-01
- 期刊:
- 影响因子:0
- 作者:Badgett, Majors J;Boyes, Barry;Orlando, Ron
- 通讯作者:Orlando, Ron
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Barry E Boyes其他文献
Barry E Boyes的其他文献
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{{ truncateString('Barry E Boyes', 18)}}的其他基金
High Resolution Superficially Porous Materials for Native Protein Separations
用于天然蛋白质分离的高分辨率表面多孔材料
- 批准号:
10255623 - 财政年份:2021
- 资助金额:
$ 21.91万 - 项目类别:
Improved Protein Separations with Superficially Porous Particles
使用表面多孔颗粒改进蛋白质分离
- 批准号:
8978832 - 财政年份:2015
- 资助金额:
$ 21.91万 - 项目类别:
Ellipsoid Particles for High Performance LC Separations
用于高性能 LC 分离的椭球颗粒
- 批准号:
8590657 - 财政年份:2013
- 资助金额:
$ 21.91万 - 项目类别:
Novel Materials and Methods for Separations of Glycopeptides and Glycans
用于分离糖肽和聚糖的新材料和方法
- 批准号:
8705538 - 财政年份:2011
- 资助金额:
$ 21.91万 - 项目类别:
Smaller Particle Fused-Core Silicas for Higher Performance Separations
较小颗粒的熔核二氧化硅可实现更高的分离性能
- 批准号:
8200510 - 财政年份:2011
- 资助金额:
$ 21.91万 - 项目类别:
Novel Materials and Methods for Separations of Glycopeptides and Glycans
用于分离糖肽和聚糖的新材料和方法
- 批准号:
8124341 - 财政年份:2011
- 资助金额:
$ 21.91万 - 项目类别:
Novel Materials and Methods for Separations of Glycopeptides and Glycans
用于分离糖肽和聚糖的新材料和方法
- 批准号:
8455999 - 财政年份:2011
- 资助金额:
$ 21.91万 - 项目类别:
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- 批准号:
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- 资助金额:
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