Potential-Resolved Electrochemiluminescence for Determination of Two Antigens at the Cell Surface

Abstract

The potential-resolved electrochemiluminescence (ECL) was achieved for the determination of two antigens at the cell surface through a potential scanning on the electrode. Luminol and Ru(bpy)3(2+) groups as ECL probes were linked with the antibodies to recognize the corresponding antigens on the cell surface. A self-quenching of luminescence from the luminol group under negative potential was initialized by the introduction of concentrated aqueous luminol, which offered accurate measurements of the luminescence from luminol and Ru(bpy)3(2+) groups under positive and negative potentials, respectively. Using this strategy, carcinoembryonic (CEA) and alphafetoprotein (AFP) antigens on cells as the models were quantified serially through a potential scanning. Different patterns of luminescence were observed at MCF 7 and PC 3 cells, which exhibited that the assay can characterize the cells with a difference expression of antigens. Compared with fluorescence measurement, the potential resolved ECL for the detection of two analytes was not limited by the spectrum difference of probes. The strategy involving potential-induced signals required a simplified optical setup and eventually offered an alternative imaging method for multiply antigens in immunohistochemistry.

Annihilation electrogenerated chemiluminescence of mixed metal chelates in solution: modulating emission colour by manipulating the energetics

“…17–19 Similarly, the co-reactant ECL of Ru(bpy) 3 2+ and peroxydisulfate has recently been used in conjunction with luminol ECL detection in a potential-resolved immunoassay of two different antigens at a cell surface. 20 …”

A novel “dual-potential” electrochemiluminescence aptasensor array using CdS quantum dots and luminol-gold nanoparticles as labels for simultaneous detection of malachite green and chloramphenicol

“…Among these methods, ECL has become one of the predominant analytical techniques because of its simple instrumentation, high sensitivity, and wide dynamic range (Feng et al, 2014). For example, Han et al (2014) have successfully constructed a potential-resolved ECL immunosensor using Ru(bpy) 3 2+ and luminol as probes for determination of two antigens at the cell surface on one electrode. Unfortunately, there is a strong ECL signals interference between Ru(bpy) 3 2+ and luminol, which can affect the accuracy of the assay results.…”

Quantum Dots Based Potential-Resolution Dual-Targets Electrochemiluminescent Immunosensor for Subtype of Tumor Marker and Its Serological Evaluation

“…Hogan's pioneer works 34,35 realized resolution of three ECL peaks from Ru and Ir complexes, by both potential-and wavelength-resolution in acetonitrile. Jiang's group 36 reported a potential-resolution based ECL system in the water phase for simultaneous detection of two proteins on the cell surface, using Ru(bpy) 3 2+ /luminol as ECL luminophores. In addition, based on ECL potential-resolution of Ru(bpy) 3 2+ /carbon nanodots as luminophores, combining with microfluidics devices and a switch for potential control, multiplexed targets were detected simultaneously by switchover of the detection potential at +1.2 V and −1.2 V. 37 These pioneer works provided a huge impulse for multiplexed target detection based on the ECL potential-resolution strategy.…”