Institute of Quantum Science and Engineering, Texas A&M University, College Station, TX 77843.
Physics Department, Baylor University, Waco, TX 76798.
Proc Natl Acad Sci U S A. 2023 Jul 25;120(30):e2218826120. doi: 10.1073/pnas.2218826120. Epub 2023 Jul 18.
Development of a simple, label-free screening technique capable of precisely and directly sensing interaction-in-solution over a size range from small molecules to large proteins such as antibodies could offer an important tool for researchers and pharmaceutical companies in the field of drug development. In this work, we present a thermostable Raman interaction profiling (TRIP) technique that facilitates low-concentration and low-dose screening of binding between protein and ligand in physiologically relevant conditions. TRIP was applied to eight protein-ligand systems, and produced reproducible high-resolution Raman measurements, which were analyzed by principal component analysis. TRIP was able to resolve time-depending binding between 2,4-dinitrophenol and transthyretin, and analyze biologically relevant SARS-CoV-2 spike-antibody interactions. Mixtures of the spike receptor-binding domain with neutralizing, nonbinding, or binding but nonneutralizing antibodies revealed distinct and reproducible Raman signals. TRIP holds promise for the future developments of high-throughput drug screening and real-time binding measurements between protein and drug.
开发一种简单、无标记的筛选技术，能够精确、直接地感应溶液中相互作用的大小范围从小分子到大蛋白如抗体，可以为药物开发领域的研究人员和制药公司提供重要工具。在这项工作中，我们提出了一种热稳定的拉曼相互作用分析（TRIP）技术，该技术可以在生理相关条件下进行低浓度和低剂量的蛋白质和配体之间结合的筛选。TRIP 被应用于 8 种蛋白质-配体体系，产生了可重复的高分辨率拉曼测量结果，并用主成分分析进行了分析。TRIP 能够解析 2,4-二硝基苯酚和转甲状腺素蛋白之间随时间变化的结合，并分析与 SARS-CoV-2 刺突抗体的生物学相关相互作用。与中和、非结合或结合但非中和抗体的刺突受体结合域的混合物显示出独特且可重复的拉曼信号。TRIP 有望为高通量药物筛选和蛋白质与药物之间的实时结合测量的未来发展提供支持。
