A highly sensitive method for detection of protein based on inhibition of Ru(bpy)32+/TPrA electrochemiluminescent system

The detection of proteins is very important in the study of biochemistry procedure, and electrochemiluminescence method is a very practical tool for the study of protein folding, structure and quantification. In this work, bovine serum albumin (BSA) and casein were found to be able to significantly quench the electrochemiluminescence (ECL) of Ru(bpy)(3)(2+)/TPrA system, based on which a highly sensitive approach for the detection of protein was proposed. Under the optimized conditions, the logarithmic plot of the inhibited ECL versus the concentration of BSA and casein were linear over the ranges of 1-30 mu g/L and 0.1-1.6 mu g/L. respectively. The corresponding limit of detection CLOD) was 0.45 mu g/L for BSA and 0.026 mu g/L for casein (S/N = 3). UV, ECL and fluorescence methods were used to investigate the mechanism of the inhibited ECL of Ru(bpy)(3)(2+)/TPrA/BSA system. A mechanism based on the formation of protein-Ru(bpy)(3)(2+) super molecule was proposed, which would prevent Ru(bpy)(3)(2+) from reaching the working electrode surface so that induced ECL quenching. (C) 2011 Elsevier Ltd. All rights reserved.