Fluorous Quantitative Proteomics

荧光定量蛋白质组学

• 批准号: 7294320
• 负责人: Marvin S Yu
• 金额: $ 12.43万
• 依托单位: FLUOROUS TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-30 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7294320
• 关键词: Address; Affect; Aniline; Area; Attention; Benchmarking; Biological; Biological Markers; Businesses; Cells; Chemicals; Chemistry; Class; Commercial Sources; Complex; Condition; Cysteine; Detection; Development; Diagnostic; Disease; Electrospray Ionization; Foundations; Gene Expression; Genomics; Goals; Grant; Human; Human Genome; Institutes; Intellectual Property; Investigation; Isotope Labeling; Isotopes; Label; Measures; Methods; Numbers; Oxidation-Reduction; Pathway interactions; Peptides; Performance; Phase; Pliability; Post-Translational Protein Processing; Proteins; Proteomics; Protocols documentation; Reagent; Recovery; Relative (related person); Research; Research Personnel; Route; Sampling; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stable Isotope Labeling; Standards of Weights and Measures; Techniques; Technology; Therapeutic; Vision; analog; base; commercialization; cost; liquid chromatography mass spectrometry; metabolomics; novel; novel diagnostics; protein function; stable isotope

DESCRIPTION (provided by applicant): The long-term goal of the project is to develop and then commercialize stable isotope reagents for quantitative proteomics using fluorous enrichment techniques. Successful demonstration in proteomics will be followed by extension into metabolomics and glycomics using a similar tagging strategy. Fluorous Technologies, Inc. (FTI) as the small business concern, and The Genomics Institute of the Novartis Research Foundation (GNF), as a collaborator, will synthesize fluorous isotope reagent tags (FLIRTs), develop protocols, benchmark vs. existing techniques, and demonstrate the FLIRT approach in the quantification of protein function. The specific aims of the grant are: 1) Synthesis of aniline based FLIRTs. Synthetic routes to five fluorous probes for chemical labeling of peptides and proteins will be synthesized based on a fluorous aniline core. A goal of 500 mg of each unlabeled compound and 150 mg of each labeled compound is targeted. 2) Synthesis of stable isotope labeled versions of the existing fluorous reagents. FLIRTs with the stable isotopes directly in the fluorous chain will be synthesized. The goal will be to obtain 100 mg of each compound. These compounds will be the direct stable isotope labeled analogs of fluorous peptide labeling reagents which FTI has previously produced. 3) Development of quantitative tagging protocols using the fluorous isotope labeled reagents and benchmarking vs. cICAT. Protocols for tagging and enrichment using FLIRTs will be developed using LC/MS with ESI detection. Quantification by FLIRT will then be benchmarked against cICAT. Performance measures will include enrichment efficiency, selectivity biases, selectivity enrichment, recovery of enriched species, and accuracy of quantitation. 4) Fluorous isotope reagent tagging (FLIRT) for relative quantification of a targeted subproteome. FLIRT technology will be applied to the quantification of specific classes of proteins within a complex sample, including the characterization of the redox state of the various cysteine residues within a protein. Successful enrichment and quantitation through a single label will demonstrate the flexibility of FLIRTs in analyzing targeted classes of proteins. FLIRTs will provide a novel method for quantitative proteomics which can be applied to numerous research areas including target discovery, biomarker discovery, and diagnostics development for various diseases affecting the human condition. The goal of the project is to develop and commercialize reagents for quantitative proteomics. The successful commercialization of these products will enable researchers to elucidate biological pathways leading to new biomarkers and targets by which to develop novel diagnostics and therapeutics for various diseases that affect the human condition.

描述（由申请人提供）：该项目的长期目标是利用荧光富集技术开发用于定量蛋白质组学的稳定同位素试剂，然后将其商业化。在蛋白质组学中的成功演示之后，将使用类似的标记策略扩展到代谢组学和糖组学。 Fluorous Technologies, Inc. (FTI) 作为小型企业，诺华研究基金会基因组研究所 (GNF) 作为合作者，将合成氟同位素试剂标签 (FLIRT)、开发方案、与现有技术进行基准比较、并展示 FLIRT 方法在蛋白质功能定量中的应用。该资助的具体目标是：1）基于苯胺的 FLIRT 的合成。用于化学标记肽和蛋白质的五种氟探针的合成路线将基于氟苯胺核心来合成。目标是每种未标记化合物 500 毫克，每种标记化合物 150 毫克。 2) 现有氟试剂的稳定同位素标记版本的合成。将合成直接在氟链中具有稳定同位素的 FLIRT。目标是获得每种化合物 100 毫克。这些化合物将是FTI先前生产的氟肽标记试剂的直接稳定同位素标记类似物。 3) 使用氟同位素标记试剂开发定量标记方案并与 cICAT 进行基准测试。使用 FLIRT 进行标记和富集的方案将使用 LC/MS 和 ESI 检测来开发。然后，FLIRT 的量化将根据 cICAT 进行基准测试。绩效衡量标准包括富集效率、选择性偏差、选择性富集、富集物种的回收率和定量准确性。 4) 氟同位素试剂标记 (FLIRT)，用于目标亚蛋白质组的相对定量。 FLIRT 技术将应用于复杂样品中特定类别蛋白质的定量，包括蛋白质内各种半胱氨酸残基氧化还原状态的表征。通过单一标签成功进行富集和定量将证明 FLIRT 在分析目标蛋白质类别方面的灵活性。 FLIRT 将为定量蛋白质组学提供一种新方法，该方法可应用于众多研究领域，包括靶点发现、生物标志物发现以及影响人类状况的各种疾病的诊断开发。该项目的目标是开发定量蛋白质组学试剂并将其商业化。这些产品的成功商业化将使研究人员能够阐明导致新生物标志物和靶标的生物途径，从而开发针对影响人类状况的各种疾病的新型诊断和治疗方法。