Im Hyungsoon, Wittenberg Nathan J, Lesuffleur Antoine, Lindquist Nathan C, Oh Sang-Hyun
Chem Sci. 2010 Jan 1;1(6):688-696. doi: 10.1039/C0SC00365D.
Integration of solid-state biosensors and lipid bilayer membranes is important for membrane protein research and drug discovery. In these sensors, it is critical that the solid-state sensing material does not have adverse effects on the conformation or functionality of membrane-bound molecules. In this work, pore-spanning lipid membranes are formed over an array of periodic nanopores in free-standing gold films for surface plasmon resonance (SPR) kinetic binding assays. The ability to perform kinetic assays with a transmembrane protein is demonstrated with α-hemolysin (α-HL). The incorporation of α-HL into the membrane followed by specific antibody binding (anti-α-HL) red-shifts the plasmon resonance of the gold nanopore array, which is optically monitored in real time. Subsequent fluorescence imaging reveals that the antibodies primarily bind in nanopore regions, indicating that α-HL incorporation preferentially occurs into areas of pore-spanning lipid membranes.
固态生物传感器与脂质双分子层膜的整合对于膜蛋白研究和药物发现至关重要。在这些传感器中，固态传感材料对膜结合分子的构象或功能没有不利影响至关重要。在这项工作中，跨膜脂质膜在独立金膜中的周期性纳米孔阵列上形成，用于表面等离子体共振（SPR）动力学结合测定。用α-溶血素（α-HL）证明了对跨膜蛋白进行动力学测定的能力。将α-HL掺入膜中，随后特异性抗体结合（抗α-HL）使金纳米孔阵列的等离子体共振发生红移，实时对其进行光学监测。随后的荧光成像显示，抗体主要结合在纳米孔区域，表明α-HL掺入优先发生在跨膜脂质膜区域。
