Id Protein Assay for Diagnosis & Drug Screening

用于诊断的 Id 蛋白测定

• 批准号: 6741681
• 负责人: JOHN CHEN
• 金额: $ 9.87万
• 依托单位: BIOCHECK, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-03-09 至 2004-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6741681
• 关键词: angiogenesis inhibitors; antineoplastics; autoradiography; biotechnology; biotherapeutic agent; breast neoplasms; diagnosis design /evaluation; drug discovery /isolation; drug screening /evaluation; enzyme linked immunosorbent assay; genetically modified animals; high throughput technology; laboratory mouse; method development; neoplasm /cancer diagnosis; prostate neoplasms; protein protein interaction; protein quantitation /detection; recombinant proteins; transcription factor

DESCRIPTION (provided by applicant): The Idl &ld3 genes and proteins are required for tumor-associated angiogenesis. We hypothesize that inhibiting the activity of the Idl and/or Id3 transcription activators will retard the growth & metastases of tumors. However, the effort to discover an orally active, small molecule, anti-cancer drug by identifying an molecule that inhibits the action of the Idl & Id3 proteins requires bioanalytical methods to screen for small molecules that inhibit the binding of Idl & Id3 to their natural ligands, the E-proteins, and also to establish which cancers would be most likely to respond to an anti-ld treatment. Such bioanalytical method are not currently available for the Id proteins, but we recently obtained promising preliminary results by monitoring the extent of the interaction of horseradish peroxidase-labeled E47 with Idl captured by an anti-ldl antibody bound to microtiter plates. This procedure resembles a traditional ELISA, but uses a natural protein ligand (E47) to the analyte (Idl) instead of a second antibody to detect capture. When fully developed and validated, this method will be used to both measure Idl & Id3 in the blood of patients with certain advanced cancers to determine whether it has potential as a diagnostic method, and also to evaluate the ability of small molecule inhibitors to reduce Id activity. To advance this effort, we plan to (1) validate the pilot assay with respect to limit of quantitation, precision, accuracy, reproducibility, repeatability and stability, (2) use the validated assay to determine if Id concentrations can be reliably measured in the serum and tissue from different mouse models of human cancer, and (3) use the validated assay, configured with fixed & physiologically relevant concentrations of Idl and E47 to explore disruption of Idl binding to E47 at various concentrations of candidate small molecule inhibitors of binding. It is expected that completion of the research described in this grant will result in an assay useful for high throughput screening of chemical libraries for anti-ld activity, and in the selection of those cancers, and perhaps stages of cancer, where anti- Id treatment is most likely to be effective. The assay would, at a minimum, have the potential to be developed into a practical diagnostic/prognostic product to select patients for clinical trials and to monitor their response to anti-ld therapy. It is also possible that the assay could be a commercial product depending on the technical and medical success achieved.

描述（由申请人提供）：IDL＆LD3基因和蛋白质是肿瘤相关血管生成所必需的。我们假设抑制IDL和/或ID3转录激活剂的活性会阻碍肿瘤的生长和转移。 However, the effort to discover an orally active, small molecule, anti-cancer drug by identifying an molecule that inhibits the action of the Idl & Id3 proteins requires bioanalytical methods to screen for small molecules that inhibit the binding of Idl & Id3 to their natural ligands, the E-proteins, and also to establish which cancers would be most likely to respond to an anti-ld treatment.此类生物分析方法目前尚不适合ID蛋白，但是我们最近通过监测荷龙过氧化物酶标记的E47与由与微滴定剂板结合的抗LDL抗体捕获的IDL的相互作用的程度，从而获得了有希望的初步结果。该过程类似于传统的ELISA，但使用天然蛋白质配体（E47）对分析物（IDL）而不是第二种抗体来检测捕获。当完全开发和验证时，该方法将用于测量患有某些晚期癌症患者血液中的IDL和ID3，以确定其是否具有诊断方法的潜力，并评估小分子抑制剂减少ID活性的能力。为了促进这项努力，我们计划（1）在定量，精确，准确性，可重复性，可重复性，可重复性和稳定性方面验证试验测定法，（2）使用经过验证的测定确定ID浓度可以可靠地在血清中可靠地测量人类癌症的不同小鼠模型，以及与固定的和（3）相关的识别，以及（3）使用固定的识别和（3），并使用固定的浓度，以实现固定的浓度，以实现固定的浓度，以实现固定的浓度，以实现固定的浓度，以实现固定的浓度，并将其概述为固定的，（3） IDL与E47的结合在各种候选候选小分子抑制剂下结合。可以预期，该赠款中描述的研究的完成将导致一种用于对抗LLD活性的化学库的高吞吐量筛查，以及选择这些癌症以及癌症治疗最有效的癌症阶段的测定法。该测定最少有可能发展为实用的诊断/预后产品，以选择患者进行临床试验，并监测其对抗LLD治疗的反应。根据技术和医疗成功，该测定法可能是商业产品。