Open AccessQuantification of Single-Molecule Protein Binding Kinetics in Complex Media with Prism-Coupled Plasmonic Scattering Imaging
Author(s) -
Pengfei Zhang,
Guangzhong Ma,
Zijian Wan,
Shaopeng Wang
Publication year - 2021
Publication title -
acs sensors
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.055
H-Index - 57
ISSN - 2379-3694
DOI - 10.1021/acssensors.0c02729
Subject(s) - surface plasmon resonance , analyte , prism , molecular binding , chemistry , molecule , total internal reflection fluorescence microscope , nanotechnology , receptor–ligand kinetics , small molecule , plasmon , materials science , nanoparticle , chromatography , optics , physics , organic chemistry , optoelectronics , biochemistry , membrane , receptor
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.