Electrochemical detection of C-reactive protein using Copper nanoparticles and hybridization chain reaction amplifying signal

Abstract

In this study, a sandwich-type electrochemical immunosensor for the detection of C-reactive protein (CRP) is described. In design, Copper nanoparticles (Cu NPs) were used for signal tag and hybridization chain reaction (HCR)amplified output signal. The immunosensor fabrication involved three steps: (i) primary antibodies (Ab1) were immobilized on the surface of gold nanoparticles (Au NPs); (ii) the sandwich-type structure formation contained “primary antibodies-antigen-secondary antibodies conjugated with primer (Ab2-S0)”; and (iii) long DNA concatemers intercalating amounts of Cu NPs was linked to the sandwich-type structure via hybridization reaction. Differential pulse voltammetry (DPV) was used to record the response signal of the immunosensor in phosphate-buffered saline (PBS). Under optimal conditions, the anodic peak currents of Cu NPs at the peak potential of about 0.08V(VS.SCE) were linear with the logarithm of CRP concentration in the range of 1.0 fg mL−1 to 100 ng mL−1 with a detection limit of 0.33 fg mL−1 (at signal/noise [S/N] = 3). In addition, the practical application of immunosensor was evaluated by analyzing CRP in real human serum samples, the recoveries obtained were within 95.3%–103.8%, indicating the immunosensor possessed potential application ability for practical disease diagnosis.