Zingale Gabriele Antonio, Pandino Irene, Calcagno Damiano, Perina Maria Luisa, Tuccitto Nunzio, Grasso Giuseppe
IRCCS-Fondazione Bietti, Rome, Italy.
Department of Chemical Sciences, University of Catania, Catania, Italy.
PLoS One. 2025 Jan 7;20(1):e0312594. doi: 10.1371/journal.pone.0312594. eCollection 2025.
Surface plasmon resonance (SPR) is normally used to measure the kinetic parameters of biomolecular interactions between a molecule immobilized on a gold surface and another one flowing in a microfluidic channel above the surface. During the SPR measurements, convection-diffusion phenomena occur inside the microfluidic channels, but they are generally minimized by appropriate experimental setup in order to obtain diffusion free kinetic parameters of the molecular interactions. In this work, for the first time, a commercial SPR apparatus has been used to obtain non canonical scientific parameters. Indeed, a specifically designed SPR experimental setup is described for carrying out measurements of the diffusion coefficient (D) of molecules in solutions. The high precision and reproducibility of the approach, as well as the wide applicability of the newly proposed SPR based method for the measurement of D of many different molecules and biomolecules, are here demonstrated and illustrated in detail.
表面等离子体共振(SPR)通常用于测量固定在金表面的分子与在该表面上方微流控通道中流动的另一个分子之间生物分子相互作用的动力学参数。在SPR测量过程中,微流控通道内会出现对流扩散现象,但通常通过适当的实验设置将其最小化,以便获得分子相互作用的无扩散动力学参数。在这项工作中,首次使用商业SPR仪器来获取非标准科学参数。实际上,描述了一种专门设计的SPR实验装置,用于测量溶液中分子的扩散系数(D)。本文详细展示并说明了该方法的高精度和可重复性,以及新提出的基于SPR的方法对许多不同分子和生物分子的D测量的广泛适用性。