Fluorescence Lifetime-Based Kinase Assay for HTS

基于荧光寿命的 HTS 激酶测定

• 批准号: 7053465
• 负责人: KIRK D HARTEL
• 金额: $ 1.45万
• 依托单位: DAKOTA 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: 2006-04-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7053465
• 关键词: chemical binding; cyclic AMP; drug discovery /isolation; fluorescence; fluorescent dye /probe; high throughput technology; ionophores; iron compounds; light scattering; phosphopeptides; phosphorylation; protein kinase; technology /technique development

DESCRIPTION (provided by applicant): Fluorescence based-assays have greatly facilitated high-throughput screening (HTS) in drug discovery. However, in doing so, they created a bottleneck relating to false positives and false negatives, which may arise due to interferences from light scattering, inner filter effects, and compound fluorescence. Unlike fluorescence intensity, a fluorescence lifetime-based measurement is resistant to light scattering and inner filter effects, and provides the selectivity needed to discriminate the target fluorescence signal from background fluorescence. In Phase I we will demonstrate a homogenous fluorescence lifetime-based kinase assay that is resistant to fluorescence interferences and does not require the use of antibodies. Overcoming fluorescence interferences will cut costs associated with secondary screening and assist quicker identification of lead compounds. Furthermore, it will create an opportunity to screen compound libraries and/or natural products that are known to have some fluorescent properties, but were previously inaccessible by means of traditional fluorescence-based methods. Kinases are a major class of drug targets, which are involved in human diseases such as cancer, diabetes, and arthritis. This technology will advance the speed and efficiency in which we can find drugs to treat these important diseases.

描述（由申请人提供）：基于荧光的分析极大地促进了药物发现中的高通量筛选（HTS）。然而，在这样做的过程中，他们创造了一个与假阳性和假阴性有关的瓶颈，这可能是由于光散射、内部过滤效应和复合荧光的干扰而产生的。与荧光强度不同，基于荧光寿命的测量可以抵抗光散射和内部滤波效应，并提供区分目标荧光信号和背景荧光所需的选择性。在第一阶段，我们将展示一种基于寿命的同质荧光激酶检测，该检测抵抗荧光干扰，不需要使用抗体。克服荧光干扰将降低与二次筛选相关的成本，并有助于更快地识别先导化合物。此外，它将为筛选已知具有某些荧光特性的化合物库和/或天然产物创造机会，这些化合物库和/或天然产物以前通过传统的基于荧光的方法无法获得。激酶是一类主要的药物靶点，与癌症、糖尿病和关节炎等人类疾病有关。这项技术将提高我们找到治疗这些重要疾病的药物的速度和效率。