Double-Layer Nanogold and Double-Strand DNA-Modified Electrode for Electrochemical Immunoassay of Cancer Antigen 15-3

Various sensor-based immunoassay methods have been extensively developed for the detection of cancer antigen 15-3 (CA 15-3), but most often exhibit low detection signals and low detection sensitivity, and are unsuitable for routine use. The aim of this work is to develop a simple and sensitive electrochemical immunoassay for CA 15-3 in human serum by using nanogold and DNA-modified immunosensors. Prussian blue (PB), as a good mediator, was initially electrodeposited on a gold electrode surface, then double-layer nanogold particles and double-strand DNA (dsDNA) with the sandwich-type architecture were constructed on the PB-modified surface in turn, and then anti-CA 15-3 antibodies were adsorbed onto the surface of nanogold particles. The double-layer nanogold particles provided a good microenvironment for the immobilization of biomolecules. The presence of dsDNA enhanced the surface coverage of protein, and improved the sensitivity of the immunosensor. The performance and factors influencing the performance of the immunosensor were evaluated. Under optimal conditions, the proposed immunosensor exhibited a wide linear range from 1.0 to 240 ng/mL with a relatively low detection limit of 0.6 ng/mL (SIN=3) towards CA 15-3. The stability, reproducibility and precision of the as-prepared immunosensor were acceptable. 57 serum specimens were assayed by the developed immunosensor and standard enzyme-linked immunosorbent assay (ELISA), respectively, and the results obtained were almost consistent. More importantly, the proposed methodology could be further developed for the immobilization of other proteins and biocompounds.