Quantification of Single-Molecule Protein Binding Kinetics in Complex Media with Prism-Coupled Plasmonic Scattering Imaging
journal contributionposted on 15.03.2021, 16:39 by Pengfei Zhang, Guangzhong Ma, Zijian Wan, Shaopeng Wang
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.
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surface plasmon resonanceComplex Mediaindividual-bound analyte moleculesbinding dynamicsscreening drugssingle-molecule imagingSingle-Molecule Protein Binding Kin...mediaunderstanding molecular-scalesingle-molecule-based measurementprism-coupled SPR systemsPrism-Coupled Plasmonic Scattering ...binding issuesKretschmann prism-coupled SPR systembinding processessurface chemistryLabel-free detection technologies