In Situ Generation of Prussian Blue with Potassium Ferrocyanide to Improve the Sensitivity of Chemiluminescence Immunoassay Using Magnetic Nanoparticles as Label.

Using magnetic nanoparticles (MNPs) as a label in immunoassay (IA) possesses advantages such as high specific surface area, simple modification process. However, the catalytic activity of MNPs is low, which limits their applications in IA. The present study found it interesting that potassium ferrocyanide reacts with MNPs, leading to the in situ generation of Prussian blue. The produced Prussian blue shows high catalytic activity on a luminol chemiluminescent (CL) reaction. Therefore, a simple and sensitive immunoassay for rabbit IgG (rIgG) as model analyte using MNPs as label was developed. The CL intensity had a linear increase with the concentration of rIgG that ranged from 0.625 to 20 ng mL. The limit of detection was calculated to be 0.59 ng mL. In addition, the applicability of this method was evaluated using the standard addition method. The recovery ranged from 80.0% to 115.0%. What's more, the proposed CLIA method based on in situ generation of Prussian blue with MNPs was also applied to the detection of carcinoembryonic antigen (CEA) and hepatitis B virus (HBV)-related sequence-specific DNA. The LOD for the detection of CEA and sequence-specific DNA was estimated to be 0.28 ng mL and 0.044 pmol, respectively.