Zhang Pengfei, Ma Guangzhong, Wan Zijian, Wang Shaopeng
Biodesign Center for Bioelectronics and Biosensors, Arizona State University, Tempe, Arizona 85287 United States.
School of Electrical, Energy and Computer Engineering, Arizona State University, Tempe, Arizona 85287, United States.
ACS Sens. 2021 Mar 26;6(3):1357-1366. doi: 10.1021/acssensors.0c02729. Epub 2021 Mar 15.
Measuring molecular binding is critical for understanding molecular-scale biological processes and screening drugs. Label-free detection technologies, such as surface plasmon resonance (SPR), have been developed for analyzing analytes in their natural forms. However, the specificity of these methods is solely relying on surface chemistry and has often nonspecific binding issues when working with samples in complex media. Herein, we show that single-molecule-based measurement can distinct specific and nonspecific binding processes by quantifying the mass and binding dynamics of individual-bound analyte molecules, thus allowing the binding kinetic analysis in complex media such as serum. In addition, this single-molecule imaging is realized in a commonly used Kretschmann prism-coupled SPR system, thus providing a convenient solution to realize high-resolution imaging on widely used prism-coupled SPR systems.
测量分子结合对于理解分子尺度的生物过程和筛选药物至关重要。无标记检测技术，如表面等离子体共振（SPR），已被开发用于分析天然形式的分析物。然而，这些方法的特异性仅依赖于表面化学，并且在处理复杂介质中的样品时经常存在非特异性结合问题。在此，我们表明基于单分子的测量可以通过量化单个结合分析物分子的质量和结合动力学来区分特异性和非特异性结合过程，从而允许在血清等复杂介质中进行结合动力学分析。此外，这种单分子成像在常用的Kretschmann棱镜耦合SPR系统中实现，从而为在广泛使用的棱镜耦合SPR系统上实现高分辨率成像提供了一种便捷的解决方案。
