Quartz Crystal Microbalance Detection of Glutathione-Protected Nanoclusters Using Antibody Recognition

Abstract

A quartz crystal microbalance (QCM) immunosensor was developed for the quantitative detection of glutathione-protected nanoclusters. Advantages intrinsic to QCM were employed to make it an attractive alternative to other immunosensing techniques. We have addressed challenges in the area of QCM mass sensing through experimental correlation between damping resistance and frequency change for a reliable mass measurement. Electrode functionalization was optimized with the use of protein A to immobilize and present polyclonal IgG for antigen binding. This method was developed for the detection of glutathione (antigen)-protected clusters of nanometer size with high surface area and thiolate valency. Quantitation of glutathione−nanocluster binding to immobilized polyclonal antibody provides equilibrium constants (K_a = (3.6 ± 0.2) × 10⁵ M^-1) and kinetic rate constants (k_f = (5.4 ± 0.7) × 10¹ M^-1 s^-1 and k_r = (1.5 ± 0.4) × 10^-4 s^-1) comparable to literature reports. These observations further imply that immunoreactive nanoparticles have potential in medical diagnostics and materials assembly.