Differential Capture Proteomics-Diabetes

描述(申请人提供)：2002年，在美国，超过2000万人患有糖尿病，导致超过22.4万人死亡，糖尿病的总成本为1320亿美元。现在预计糖尿病将在世界范围内流行，这将产生严重的医疗和经济后果。新生物标志物的发现将对这些破坏性疾病的筛查、诊断、预后、疾病复发预测和治疗监测方面的迫切需要的进展非常重要。 该项目第一阶段的目标是在人类血浆中优化一种新的蛋白质组学技术-差异捕获蛋白质组学(DCP)，该技术可以识别两个生物样本之间的蛋白质组成差异。这项技术还可以同时为每个已识别的差异蛋白创建亲和试剂。DCP的专利已经颁发给差异蛋白质组学公司。 后续第二阶段研究的目标是将最近针对人类血浆进行优化的DCP应用于发现糖尿病的生物标记物，使用适当的患者血浆样本，以实现新的蛋白质分离和鉴定、新的亲和试剂的创建、分析配方和临床验证。 我们实验室对DCP技术的优化研究已经成功地集中在一个模型系统上。然而，DCP在患者血浆样本中发现糖尿病生物标记物的能力尚未得到优化研究的证实。这里提出了这样一项研究，其中将测试掺杂的糖尿病生物标记物(代表模型糖尿病患者样本)在临床样本水平浓度下的可检测性。这项拟议的研究将使用五种特定的糖尿病蛋白抗原，对于这些抗原，有商业上可用的免疫分析。这项第一阶段研究的成功将导致第二阶段计划，测试多个糖尿病患者血浆样本，以发现、分析和验证新的生物标记物。 具体目标1：确认三个随机肽噬菌体文库实际上包含针对所选择的五种糖尿病抗原的结合噬菌体物种。具体目标2：展示差异捕获蛋白质组学如何很好地检测两个人类血浆样本之间的差异，其中一个样本掺杂了五种糖尿病抗原的混合物。具体目标3：确定作为该过程一部分产生的亲和捕获试剂的特异性。

DESCRIPTION (provided by applicant): In the United States in 2002, over 20 million people had diabetes, which contributed to over 224,000 deaths and the total cost of diabetes was $132 billion. A worldwide epidemic of diabetes is now projected which will have serious healthcare and economic consequences. The discovery of new biomarkers will be very important for much-needed advances in screening, diagnosis, prognosis, prediction of disease recurrence and therapeutic monitoring for these devastating diseases. The goal for Phase I of this Project is to optimize in human plasma a new proteomics technology, Differential Capture Proteomics (DCP), which can identify the differences in protein composition between two biological samples. This technology also simultaneously creates affinity reagents for each of the identified difference proteins. A patent for DCP has been issued to Differential Proteomics Inc. The goal for a subsequent Phase II study is to apply DCP, newly optimized for human plasma, to the discovery of biomarkers of diabetes using appropriate patient plasma samples to enable novel protein isolation and identification, novel affinity reagent creation, assay formulation, and clinical validation. Studies in our laboratory to optimize the DCP technology have successfully focused on a model system. However, the capabilities of DCP for diabetes biomarker discovery in patient plasma samples have yet to be demonstrated with an optimization study. Such a study is proposed here, in which doped diabetes biomarkers (representing a model diabetes patient sample) will be tested for detectability at clinical sample level concentrations. The proposed study will use five specific diabetes protein antigens, for which there are commercially available immunoassays. The success of this Phase I study will lead to a Phase II program for testing multiple diabetes patient plasma samples for novel biomarker discovery, assay generation, and validation. Specific Aim 1: Confirm that three pooled Random Peptide Phage Libraries actually contain binding phage species against each of the five diabetes antigens chosen. Specific Aim 2: Show how well Differential Capture Proteomics detects the differences between two human plasma samples, one of which is doped with the mixture of five diabetes antigens. Specific Aim 3: Establish the specificity of the affinity capture reagents that are generated as part of the process.