Immunoglobulin G-Encapsulated Gold Nanoclusters as Fluorescent Tags for Dot-Blot Immunoassays.

Abstract

Few-atom gold nanoclusters (AuNCs) have been fabricated and used for various fields owing to their remarkable optical and photophysical features. However, the rational design for the antibody-mediated synthesis of fluorescent AuNCs for direct antigen-antibody reactions remains unexplored. In this work, immunoglobulin G (IgG)-functionalized AuNCs (IgG-AuNCs) were successfully prepared via a facile and fast biomineralization process. The generated IgG-AuNCs can emit intense red fluorescence with a high photoluminescence quantum yield. Besides strong emission, the bioactivity of IgG on the IgG-AuNCs can be retained. Surface plasmon resonance measurements suggested that IgG-AuNCs can bind to goat anti-human IgG with an affinity constant of 6.21 × 10-8 M. A simple detection method was then developed using a dot-blot immunoassay with IgG-AuNCs as fluorescent tags. Experimental results confirmed that the IgG-AuNC-based fluorescent reporters had many advantages such as low nonspecific adsorption and good photostability, offering immense potential for the development of efficient biosensors. This work can be extended to other specific antibodies to produce multifunctional AuNCs and utilized to detect and monitor targeted analytes and biological events of interest.