We report an online ligand screening method that targets human glucose transporter 1 (hGlut1) under approximately physiological conditions, named nonimmobilized biomaterial capillary electrophoresis (NIBCE), and we investigated the interactions between drugs/candidate compounds and HEK293 cells, hGlut1-overexpressing HEK293 cells, non-small-cell lung cancer A549 cells, A549 tumor tissue, and normal lung tissue by simulating the interactions between drugs and moving target cells or the space-occupying tumor. NIBCE omits the trouble of isolating and purifying target receptors from cell membrane while maintaining their native conformation and binding activity. The biomaterials were intercepted by porous frits in capillary columns and cannot flow through the detection window, thereby solving the problem of interference detection, and they can be renewed any time flexibly, thus effectively maintaining their surface bioactivity. Furthermore, the binding kinetic parameters (K, k_a, k_d, and k′) were calculated by nonlinear chromatography (NLC) theory, and competitive binding experiments, ligand docking studies, and antitumor activity assays in vitro and in vivo were performed to verify the feasibility of NIBCE.

中文翻译：

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非固定化生物材料毛细管电泳用于筛选靶向人葡萄糖转运蛋白1的药物

我们报告了一种在线配体筛选方法，该方法在近似生理条件下靶向人葡萄糖转运蛋白1（hGlut1），称为非固定化生物材料毛细管电泳（NIBCE），并且我们研究了药物/候选化合物与HEK293细胞，hGlut1过表达的HEK293细胞之间的相互作用，非小细胞肺癌A549细胞，A549肿瘤组织和正常肺组织，通过模拟药物与移动靶细胞或占位肿瘤之间的相互作用来实现。NIBCE省去了从细胞膜分离和纯化靶受体同时保持其天然构象和结合​​活性的麻烦。生物材料被毛细管柱中的多孔熔块拦截，无法通过检测窗口，从而解决了干扰检测的问题，并且可以随时灵活地更新它们，从而有效地保持其表面生物活性。此外，结合动力学参数（通过非线性色谱（NLC）理论计算K，k _a，k _d和k ′），并进行了体外和体内竞争性结合实验，配体对接研究和抗肿瘤活性试验，以验证NIBCE的可行性。