Greenway Harmon, Wang Jin
The Verna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of Medicine, Houston, Texas 77030, United States.
Center for NextGen Therapeutics, Baylor College of Medicine, Houston, Texas 77030, United States.
ACS Med Chem Lett. 2024 Aug 30;15(9):1598-1605. doi: 10.1021/acsmedchemlett.4c00317. eCollection 2024 Sep 12.
The rapid growth of therapeutic monoclonal antibodies demands greater accessibility to scalable methods of evaluating antigen binding. Homogenous TR-FRET is ideal for preliminary screening but has not been reported to assay these interactions due to their high-affinity and complex solution-phase kinetics. Here we report the development of a competition assay to rank-order the relative affinities of these drugs for a common antigen. The assay is compatible with automation, requires no modification of the analytes, and measures affinities as low as single-digit picomolar. We further demonstrate applications to inform the development of antibody-drug conjugates. The assay may aid discovery and manufacturing of therapeutic antibodies as a low-cost, high-throughput alternative to existing technologies.
治疗性单克隆抗体的快速增长要求更易于获得可扩展的抗原结合评估方法。均相时间分辨荧光能量共振转移（TR-FRET）非常适合初步筛选，但由于这些相互作用具有高亲和力和复杂的溶液相动力学，尚未见有报道用于检测这些相互作用。在此，我们报告了一种竞争分析方法的开发，用于对这些药物与共同抗原的相对亲和力进行排序。该分析方法与自动化兼容，无需对分析物进行修饰，并且能够测量低至个位数皮摩尔的亲和力。我们进一步展示了其在指导抗体-药物偶联物开发方面的应用。该分析方法可能有助于治疗性抗体的发现和生产，作为现有技术的一种低成本、高通量替代方法。
