High-Throughput Screening for the Discovery of Protein-Binding Fragments

用于发现蛋白质结合片段的高通量筛选

• 批准号: 10759032
• 负责人: Weijun Gui
• 金额: $ 27.49万
• 依托单位: DELUGE BIOTECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10759032
• 关键词: Acylation; Affinity; Avidity; Binding; Binding Proteins; Chimeric Proteins; Color; Complex; DNA; Derivation procedure; Development; Dyes; Equilibrium; Event; Flow Cytometry; Fluorescence; Fluorescent Dyes; Glues; Goals; Hydrophobicity; Incubated; Isotope Labeling; Label; Lead; Libraries; Ligands; Link; Measures; Methods; Molecular; Molecular Weight; Monitor; Outcome; Phase; Plant Resins; Population; Protein Fragment; Proteins; Scheme; Source; Streptavidin; Surface; System; Techniques; Technology; Tentagel resin; Testing; Time; Tube; biophysical techniques; dalton; drug development; drug discovery; drug quality; experimental study; high throughput screening; hydrophilicity; instrument; interest; meter; novel; programs; protein complex; recruit; screening; ubiquitin-protein ligase

Project Summary Fragment based drug discovery programs begin with the identification of low molecular compounds that bind weakly to the protein of interest (POI). POI-binding fragments are then elaborated and/or linked together to provide lead compounds. Current methods for screening fragment libraries have low throughput and many require large amounts of the POI, often in isotopically labeled form. This proposal will establish a novel screening platform capable of analyzing hundreds of potential fragment-protein interactions in a single tube. We will create dozens of fluorescently encoded one bead one compound (OBOC) libraries comprised of hundreds of low molecular weight compounds. They will be screened against a fluorescently labeled, tetrameric form of the POI. Avidity effects will stabilize the weak interactions sufficiently to allow the beads to be analyzed by flow cytometry, identifying those capable of retaining the POI. If necessary, proximity labeling will be explored to increase the sensitivity of the system. This technology will be employed to discover novel fragments that engage the VHL E3 Ubiquitin ligase.

项目摘要 基于片段的药物发现程序开始与鉴定低分子量的 与感兴趣的蛋白质（POI）弱结合的化合物。POI结合片段是 然后加工和/或连接在一起以提供先导化合物。的当前方法 筛选片段文库具有低通量，并且许多需要大量的 POI，通常以同位素标记的形式。该提案将建立一个新的筛选机制， 该平台能够分析数百种潜在的片段-蛋白质相互作用， 单管。我们会制造出几十个荧光编码的一种珠子一种化合物 OBOC文库包含数百种低分子量化合物。他们将 针对荧光标记的POI的四聚体形式进行筛选。亲合力效应 将使弱相互作用充分稳定， 流式细胞术，鉴定那些能够保留POI的细胞。如有必要， 将探索标记以增加系统的灵敏度。这项技术将 用于发现接合VHL E3泛素连接酶的新片段。