Single gold nanoparticle localized surface plasmon resonance spectral imaging for quantifying binding constant of carbohydrate-protein interaction.

Quantifying carbohydrate-protein (ligand-receptor) interactions is important to understand diverse biological processes and to develop new diagnostic and therapeutic methods. We develop an approach to quantitatively study carbohydrate-protein interactions by Au nanoparticle localized surface plasmon resonance (LSPR) peak position shift at the single particles level. Unlike the previous techniques for single particle LSPR spectral imaging, only the first-order streak of an individual nanoparticle is needed to extract a LSPR spectrum, which has great potential to increase throughput to 500 single particle spectra in each frame. LSPR peak shift of protein modified single Au nanoparticles is found to be a function of its ligand concentration, which can be used to fit the binding constants of the interactions. The moderate interactions of Antithrombin III (AT III) and heparins including low molecular weight heparin (LMWH) are determined as well as the strong interaction of transferrin and antitransferrin and the weak interaction of bovine serum album (BSA) and heparin. The measured binding constants of transferrin to antitransferrin, heparin and LMWH to AT III, and BSA to heparin are (3.0 ± 0.6) × 10(9) M(-1), (3.1 ± 0.3) × 10(6) M(-1), (8.0 ± 0.5) × 10(5) M(-1), and (5.1 ± 0.1) × 10(3) M(-1), respectively, which are in good agreement with the reported values.