Charge sensitive optical detection for high-throughput study of small molecules

项目摘要 一种可以检测和量化小分子结合和生化反应动力学的技术， 如翻译后蛋白质修饰，对于理解癌症的潜在机制至关重要 起始和进展，用于发现癌症生物标志物和用于筛选癌症候选药物。 目前，最广泛使用的技术使用荧光标记，这对于具有荧光标记的小分子来说是困难的。 尺寸与荧光染料相当，特别是对于定量动力学信息。各种无标签 技术已经开发出来，但它们的灵敏度随着分子量的增加而降低， 检测小分子和涉及小质量变化的生化反应具有挑战性。解决 基于这一需要，本课题将开发一种电荷敏感光学检测技术（CSOD）。CSOD是 与标准微孔板技术兼容，使其对高通量筛选具有吸引力， 分子相互作用和反应的分析。基本原则是在一个 IMAT（Innovative Molecular Analysis Technologies for Cancer Research）R21资助。在目前的R33项目中， 该团队将与学术研究实验室和制药公司的合作者密切合作， 生物分析仪器公司开发和验证未满足需求的技术。 该项目的成功将导致一种新的无标记高通量筛选技术， 测量分子相互作用，特别是小分子相互作用和翻译后修饰。 这些过程对于癌症的研究、诊断和治疗非常重要。

PROJECT SUMMARY A technology that can detect and quantify the kinetics of small molecule binding and biochemical reactions, such as post-translational protein modifications, is critical for understanding the mechanisms underlying cancer initiation and progression, for discovering cancer biomarkers, and for screening cancer drug candidates. Currently, the most widely used technology uses fluorescence labels, which is difficult for small molecules with sizes comparable to the fluorescent dyes, especially for quantitative kinetic information. Various label-free techniques have been developed, but their sensitivity diminishes with the molecular mass, making it extremely challenging to detect small molecules and biochemical reactions that involve small mass changes. To address this need, a charge sensitive optical detection (CSOD) technology will be developed in this project. CSOD is compatible with the standard microplate technology, making it attractive for high-throughput screening and analysis of molecular interactions and reactions. The basic principle was established with the support of an IMAT (Innovative Molecular Analysis Technologies for cancer research) R21 grant. In the present R33 project, the team will work closely with collaborators in academic research labs and pharmaceutical companies, and a bioanalytical instrument company to develop and validate the technology for the unmet need. The success of the project will lead to a new label-free high-throughput screening technology for measuring molecular interactions, particularly small molecule interactions and post-translational modifications. These processes are highly important for the research, diagnosis and treatment of cancer.