Quantitative screening for potential drug–protein binding is an essential step in developing novel metal-based anticancer drugs. ICP–MS approaches are at the core of this task; however, many applications lack in the capability of large-scale high-throughput screenings and proper validation. In this work, we critically discuss the analytical figures of merit and the potential method-based quantitative differences applying four different ICP–MS strategies to ex vivo drug–serum incubations. Two candidate drugs, more specifically, two Pt(IV) complexes with known differences of binding affinity towards serum proteins were selected. The study integrated centrifugal ultrafiltration followed by flow injection analysis, turbulent flow chromatography (TFC), and size exclusion chromatography (SEC), all combined with inductively coupled plasma-mass spectrometry (ICP–MS). As a novelty, for the first time, UHPLC SEC-ICP–MS was implemented to enable rapid protein separation to be performed within a few minutes at > 90% column recovery for protein adducts and small molecules.

中文翻译：

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对金属药物-蛋白质关联定量测量的不同方法的严格评估


定量筛选潜在的药物-蛋白质结合是开发新型金属抗癌药物的重要步骤。 ICP-MS 方法是这项任务的核心；然而，许多应用缺乏大规模高通量筛选和适当验证的能力。在这项工作中，我们批判性地讨论了分析品质因数和潜在的基于方法的定量差异，将四种不同的 ICP-MS 策略应用于离体药物-血清孵育。选择了两种候选药物，更具体地说，两种对血清蛋白的结合亲和力已知差异的 Pt(IV) 复合物。该研究整合了离心超滤、流动注射分析、湍流色谱 (TFC) 和尺寸排阻色谱 (SEC)，所有这些均与电感耦合等离子体质谱 (ICP-MS) 相结合。作为一项创新，首次采用 UHPLC SEC-ICP-MS，能够在几分钟内快速分离蛋白质，蛋白质加合物和小分子的柱回收率为 > 90%。