改进的快速平衡透析-质谱 (RED-MS) 方法用于测量小分子-蛋白质复合体在溶液中的结合亲和关系
Bryan Choi1, Calvin Han1, Jonathan R LaRochelle1
1Relay Therapeutics, Inc., Cambridge, Massachusetts 02139, United States.
Journal of the American Society for Mass Spectrometry
|November 4, 2024
概括
快速平衡透析与质谱学 (RED-MS) 结合,提供了一种可靠的方法来测量小分子与蛋白质的结合亲和力. 这种技术提高了药物发现的吞吐量,特别是对于具有挑战性的蛋白质标.
科学领域:
- 生物化学 生物化学
- 药理学 药理学是指药理学的学科.
- 分析化学 分析化学
背景情况:
- 快速平衡透析 (RED) 是评估药物吸收,分布,新陈代谢和分泌 (ADME) 特性的一种标准技术.
- 描述小分子与蛋白质的结合亲和关系对于药物发现和开发至关重要.
- 对于大型蛋白质复合体或分析众多化合物时,传统方法可能具有挑战性.
研究的目的:
- 引入和验证一种改进的快速平衡透析方法与质谱学 (RED-MS) 相结合.
- 为了使小分子与重组蛋白质和复合物结合的亲和关系能够进行稳健和高通量测量.
- 为了证明RED-MS对难以获得毒品的目标的实用性.
主要方法:
- 快速平衡透析 (RED) 与质谱学 (MS) 的整合,用于结合亲和度测量.
- 使用单个透析数据集进行亲和度计算.
- 采用复合聚合和自动化液体处理来增加测试吞吐量.
主要成果:
- RED-MS准确地确定了结合亲和力,显示了与表面等离子体共振 (SPR) 和亲和选择质谱法 (AS-MS) 强烈的相关性.
- 该方法被证明是有效的量化小分子与大型蛋白质复合体的结合,不适合其他技术.
- 实现了每周数百次测量的高吞吐量分析.
结论:
- RED-MS为测量溶液中的化合物结合提供了可靠的解决方案.
- 这种技术促进了小分子亲和力优化,特别是对于具有挑战性的蛋白质标.
- RED-MS提高了药物发现研究的效率和吞吐量.
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