Wu Hung-Jen, Singla Akshi, Weatherston Joshua D
Department of Chemical Engineering, Texas A&M University, College Station, TX, USA.
Methods Mol Biol. 2022;2460:45-63. doi: 10.1007/978-1-0716-2148-6_4.
The nature of cell membrane fluidity permits glycans, which are attached to membrane proteins and lipids, to freely diffuse on cell surfaces. Through such two-dimensional motion, some weakly binding glycans can participate in lectin binding processes, eventually changing lectin binding behaviors. This chapter discusses a plasmonic nanocube sensor that allows users to detect lectin binding kinetics in a cell membrane mimicking environment. This assay only requires standard laboratory spectrometers, including microplate readers. We describe the basics of the technology in detail, including sensor fabrication, sensor calibration, data processing, a general protocol for detecting lectin-glycan interactions, and a troubleshooting guide.
细胞膜流动性的特性使附着于膜蛋白和脂质上的聚糖能够在细胞表面自由扩散。通过这种二维运动,一些弱结合聚糖可参与凝集素结合过程,最终改变凝集素的结合行为。本章讨论了一种等离子体纳米立方体传感器,该传感器可让用户在模拟细胞膜的环境中检测凝集素结合动力学。此检测仅需标准实验室光谱仪,包括微孔板读数器。我们详细描述了该技术的基础知识,包括传感器制造、传感器校准、数据处理、检测凝集素-聚糖相互作用的一般方案以及故障排除指南。