Nonimmobilized Biomaterial Capillary Electrophoresis for Screening Drugs Targeting Human Glucose Transporter 1.

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, ka, kd, 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.