高通量SPR方法的应用和交叉验证,用于表征共价结合联体的联体
Wei Zhou1, Xuecheng Ye1, Suraj Pandey1
1Hansoh Bio, LLC, 9900 Medical Center Drive, Suite 200, Rockville, Maryland 20850, United States.
ACS omega
|July 21, 2025
概括
我们验证了一种高通量表面等离子体共振 (SPR) 方法,以快速确定抑制剂功率 (k_inact/K_I). 这种SPR方法提供了一个准确的,具有成本效益的替代方案,用于对抑制剂的传统测定.
科学领域:
- 生物化学 生化学
- 化学生物学 化学生物学
- 药物发现 药物发现 药物发现
背景情况:
- 不可逆转的共价抑制剂在药物开发中至关重要.
- 准确地确定抑制常数 (k_inact/K_I) 对于这些抑制剂的表征至关重要.
- 确定k_inact/K_I的传统方法可能耗时且昂贵.
研究的目的:
- 验证和介绍一种高通量表面等离子体共振 (SPR) 方法,用于快速确定k_inact/K_I.
- 用MK2和KRAS-G12C的共价抑制剂作为案例研究来证明SPR的实用性.
- 突出SPR相对于动力分析传统方法的优势.
主要方法:
- 使用了带有可再生传感器芯片的高通量表面等离子体共振 (SPR) 系统.
- 在方法验证中使用向MK2和KRAS-G12C的共价抑制剂.
- 将SPR衍生的k_inact/K_I值与从质谱和酶分析中获得的值进行比较.
主要成果:
- SPR 方法与 k_inact/K_I 测定的常规方法 (质谱,酶分析) 呈现出良好的相关性.
- 使用可再生生物芯片显著提高了吞吐量,降低了成本.
- SPR为不可逆转的共价抑制剂提供了准确可靠的动力学数据.
结论:
- SPR是一种高效,准确和具有成本效益的方法,用于确定k_inact/K_I.
- 在动力分析中,SPR比传统的酶和MS基方法具有显著的优势.
- 这种经过验证的SPR方法有助于在药物发现计划中快速表征共价抑制剂.
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