Combinatorial chemistry based approach to proteome exploration.

基于组合化学的蛋白质组探索方法。

• 批准号: 7110627
• 负责人: DAVID J HAMMOND
• 金额: $ 10.05万
• 依托单位: PROLIAS, LLC
• 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 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7110627
• 关键词: affinity labeling; bioassay; biomarker; combinatorial chemistry; early diagnosis; flow cytometry; fluorescent dye /probe; ligands; mass spectrometry; matrix assisted laser desorption ionization; myocardial infarction; peptide library; plasma; protein quantitation /detection; protein structure; proteomics; technology /technique development; troponin

DESCRIPTION (provided by applicant): We are developing a powerful new and proprietary approach for identification of disease-specific biomarkers in human proteomes. The basis of this approach is the balancing of proteome protein profiles using Solid-phase Peptide Combinatorial Library (SPCL) technology. The SPCL efficiently overcomes the major problem facing proteomics by decreasing the range of protein concentration without reducing the complexity. Specifically, it decreases the concentration of abundant proteins and increases the concentration of trace proteins within a sample while maintaining the difference in concentration of biomarkers between samples. Our technology comprises: A. balancing of protein concentrations in a sample using a solid-phase combinatorial peptide library (SPCL1) and eluting under non-denaturing conditions; B. differential labeling of disease state and normal balanced proteins with fluorophores; C. combining the labeled proteomes and incubating with SPCL2 for affinity separation; D. identifing and separating of beads enriched with disease state specific proteins by FACS; E. identifying bead associated proteins by MS; F. identifying peptide ligands responsible for high affinity binding of protein We will perform proof-of-principle studies to detect cardiac troponin spiked at 1 - 100 ng/ml into plasma. Assuming a sensitivity of troponin detection of < 10 ng/ml, we will analyze plasma from AMI-positive patients taken at 6 hours-post AMI for troponin and other potential AMI-related biomarkers. We expect that our technology will reveal novel biomarkers that may have significant medical importance and commercial value. The specific aims are: 1) the optimization of the individual steps of the technology; 2) the demonstration of its feasibility by detecting cardiac troponin from both spiked normal and AMI patient samples; 3) the identification of high affinity peptide ligands for troponin. These ligands would be available for evaluation of their ability to improve sensitivity of early detection of troponin in clinical assays. This technology is applicable to a wide range of other diseases, as well as for profiling patients for response to therapies. We will generate revenues by service, partnerships with companies in clinical trials, and through licensing our biomarker candidates at an advanced stage of development.

描述（由申请人提供）：我们正在开发一种强大的新的专有方法，用于鉴定人类蛋白质组中的疾病特异性生物标志物。这种方法的基础是使用固相肽组合文库（SPCL）技术平衡蛋白质组蛋白质谱。SPCL有效地克服了蛋白质组学面临的主要问题，通过减少蛋白质浓度的范围，而不降低复杂性。具体而言，它降低了样品中丰富蛋白质的浓度并增加了痕量蛋白质的浓度，同时保持了样品之间生物标志物浓度的差异。我们的技术包括：A.使用固相组合肽文库（SPCL 1）并在非变性条件下洗脱来平衡样品中的蛋白质浓度; B.用荧光团对疾病状态和正常平衡蛋白质进行差异标记; C.合并标记的蛋白质组并与SPCL 2孵育用于亲和分离; D.通过FACS鉴定和分离富含疾病状态特异性蛋白的珠粒; E.通过MS鉴定珠相关蛋白; F.鉴定负责蛋白质高亲和力结合的肽配体我们将进行原理验证研究以检测以1 - 100 ng/ml掺入血浆中的心肌肌钙蛋白。假设肌钙蛋白检测的灵敏度< 10 ng/ml，我们将分析AMI后6小时采集的AMI阳性患者血浆中的肌钙蛋白和其他潜在的AMI相关生物标志物。我们希望我们的技术将揭示可能具有重要医学意义和商业价值的新型生物标志物。具体目标是：1）对该技术的各个步骤进行优化; 2）通过检测来自加标正常和AMI患者样本的心肌肌钙蛋白来证明其可行性; 3）鉴定肌钙蛋白的高亲和力肽配体。这些配体将可用于评价它们在临床测定中提高早期检测肌钙蛋白的灵敏度的能力。这项技术适用于广泛的其他疾病，以及分析患者对治疗的反应。我们将通过服务、与临床试验中的公司合作以及在开发的高级阶段许可我们的生物标志物候选物来创收。