Department of Small Molecule Therapeutic Discovery and Research Technologies, Amgen Research, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
Department of Small Molecule Therapeutic Discovery and Research Technologies, Amgen Research, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA, 91320, USA.
Anal Biochem. 2025 Jan;696:115679. doi: 10.1016/j.ab.2024.115679. Epub 2024 Sep 26.
Binding kinetics of drug and its target protein is crucial for the efficacy and safety of the drug. Using surface plasmon resonance (SPR) technology, we performed a competitive SPR chaser assay, a method to study biomolecular interactions with very slow dissociation rate constants (k < 1E-4 s). This report described the principle and the experimental setup of the chaser assay, which involves using a competitive probe (chaser) to detect changes in target occupancy by a test molecule over time. We demonstrated the applicability of the chaser assay for both small and large molecules and compared the results with conventional SPR kinetic analysis and other methods. We suggest that the chaser assay is a useful and robust technique to characterize very tight biomolecular interactions, and that it can also be used to study cooperativity in ternary complex formation.
药物与其靶蛋白的结合动力学对于药物的疗效和安全性至关重要。我们使用表面等离子体共振（SPR）技术进行了竞争性 SPR 追赶分析，这是一种研究具有非常慢解离速率常数（k < 1E-4 s）的生物分子相互作用的方法。本报告介绍了追赶分析的原理和实验设置，该方法涉及使用竞争性探针（追赶物）来随时间检测测试分子对靶标占有率的变化。我们证明了追赶分析对小分子和大分子均适用，并将结果与传统 SPR 动力学分析和其他方法进行了比较。我们建议追赶分析是一种有用且强大的技术，可用于表征非常紧密的生物分子相互作用，并且还可用于研究三元复合物形成中的协同作用。
