Combinatorial chemistry based approach to proteome exploration.

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来识别人类蛋白质组中的疾病特异性生物标记物。该方法的基础是利用固相肽组合文库(SPCL)技术平衡蛋白质组蛋白质谱。SPCL通过在不降低复杂性的情况下减小蛋白质浓度范围，有效地克服了蛋白质组学面临的主要问题。具体地说，它降低了样品中丰富蛋白质的浓度，增加了样品中微量蛋白质的浓度，同时保持了样品之间生物标记物的浓度差异。我们的技术包括：a.使用固相组合多肽库(SPCL1)平衡样品中的蛋白质浓度并在非变性条件下洗脱；b.用荧光团对疾病状态和正常平衡的蛋白质进行差异标记；c.结合标记的蛋白质组并与SPCL2温育进行亲和分离；D.通过FACS鉴定和分离富含疾病状态特定蛋白的珠子；E.通过MS鉴定珠状相关蛋白；F.鉴定负责蛋白质高亲和性结合的多肽配体我们将进行原理验证研究，以检测血浆中添加1-100 ng/ml的心肌肌钙蛋白。假设肌钙蛋白检测的灵敏度为10 ng/ml，我们将分析急性心肌梗死阳性患者在急性心肌梗死后6小时采集的血浆中的肌钙蛋白和其他潜在的与急性心肌梗死相关的生物标志物。我们预计，我们的技术将揭示可能具有重大医学意义和商业价值的新型生物标记物。具体目标是：1)优化技术的各个步骤；2)通过检测添加正常和急性心肌梗死患者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多肽配体。这些配体将可用于评估它们在临床检测中提高肌钙蛋白早期检测的敏感性的能力。这项技术适用于广泛的其他疾病，以及患者对治疗的反应情况。我们将通过服务、与临床试验公司的合作伙伴关系以及在开发的高级阶段许可我们的生物标记物候选对象来创造收入。

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.