Luminol/antibody labeled gold nanoparticles for chemiluminescence immunoassay of carcinoembryonic antigen Original Research Article

A facile strategy by loading luminol and secondary antibody on gold nanoparticles (Au NPs) was described in the present work. The as-prepared luminol/antibody labeled Au NPs conjugates (LAAu NPs) were used as the chemiluminescent probe for the detection of carcinoembryonic antigen (CEA) in serum. The LAAu NPs were characterized by transmission electron microscopy (TEM), UV–vis spectrophotometry (UV–vis), and chemiluminescent method. Stable and efficient chemiluminescence (CL) was obtained when luminol molecules and secondary antibodies were coimmobilized on the Au NPs by using hydrogen peroxide (H2O2) as an oxidant, horseradish peroxidase (HRP) as a catalyst, and 4-(4′-iodo)phenylphenol (IPP) as an enhancer. The LAAu NPs were further evaluated via a sandwich-type CL immunoassay of CEA in serum. In this protocol, the CEA analyte was captured by the primary antibody immobilized on the surface of magnetic beads, and then was sandwiched by the secondary antibody loaded on luminol-labeled Au NPs. The chemiluminescent intensity was proportional to the concentration of CEA over the range of 5.0 × 10−10 to 5.0 × 10−8 g mL−1 and 5.0 × 10−9 to 2.0 × 10−8 g mL−1 by using HRP and Co2+ as catalysts, respectively. The present chemiluminescent immunoassay based on the luminol/antibody labeled Au NPs conjugates has offered great promise for simple, highly biocompatible, and cost-effective analysis of biological samples.