Metal-to-ligand charge-transfer: Applications to visual detection of β-galactosidase activity and sandwich immunoassay

Abstract

In this work, we report a novel use of the distinctive metal-to-ligand charge-transfer (MLCT) absorption properties of the chromogenic Fe(BPDS)34- (BPDS=bathophenanthroline disulfonic acid) reporter for the visual detection of β-galactosidase (β-Gal) activity and sandwich immunoassay. The enzymatic hydrolysis of the substrate p-aminophenyl-β-D-galactopyranoside can switch on the reduction of Fe3+ to Fe2+ and the subsequent complexation of Fe2+ with the BPDS ligand to generate the Fe(BPDS)34- reporter, leading to the appearance of the intense MLCT absorption band at 535nm and the colorless-to-red color change of the solution. Simply through a single step, the activity of β-Gal can be sensitively and selectively detected within the dynamic range of 0−220mUmL−1, with a limit of detection (LOD) of 1.69mUmL−1. This approach is applicable for the visual detection of β-Gal activities in the presence of complex human serum samples. Besides, when integrated with the sandwich immunoassay of carcinoembryonic antigen, a LOD of 1.16ngmL−1 can be achieved. In light of its prominent simplicity and practicality, our MLCT-based approach holds great potential in diagnostic and analytical applications.