Visual Sandwich Immunoassay System on the Basis of Plasmon Resonance Scattering Signals of Silver Nanoparticles

In this contribution, we established a sandwich immunoassay system with a common spectrofluorometer to collect the plasmon resonance scattering (PRS) signals from silver nanoparticles (AgNPs) immunotargeted on glass slides. By taking the immunoreactions of goat antihuman IgG (Fc fragment specific) antibody (GAH-IgG), human immunoglobulin (H-IgG), and rabbit antihuman IgG (Fab fragment specific) antibody (RAH-IgG) as an example, we found that if a primary antibody (GAH-IgG) was first immobilized on the surface of glass slides and applied to capture target antigen (H-IgG), AgNPs-labeled secondary antibody (RAH-IgG) could be employed to detect the target antigen (H-IgG) by forming a sandwich immune complex on the surface of the glass slide. It was found that the PRS signals resulting from the AgNPs immunotargeted on the glass slides could be applied to the quantitative detection of H-IgG target antigen in the range of 10-1000 ng/mL with the limit of determination of 1.46 ng/mL (3 sigma) under optimal conditions, which is sensitive and comparable with reported chemiluminescence immunoassays. With a dark-field microscope coupled with a spectral system, we measured the PRS features of single AgNPs immunotargeted on the glass slides, showing that the PRS of single nanoparticles might have potential applications in analytical chemistry. Further findings showed that the strong PRS signals from the AgNPs immunotargeted on the glass slides can be clearly seen and distinguished by naked eyes under the excitation of a common white light-emitting diode (LED) torch. Therefore, a visual PRS immunoassay system can be established easily with common glass slides and an LED torch.