Sensitive competitive flow injection chemiluminescence immunoassay for IgG using gold nanoparticle as label

A sensitive competitive flow injection chemiluminescence (CL-FIA) immunoassay for immunoglobulin G (IgG) was developed using gold nanoparticle as CL label. In the configuration, anti-IgG antibody was immobilized on a glass capillary column surface by 3-(aminopropyl)-triethoxysilane and glutaraldehyde to form immunoaffinity column. Analyte IgG and gold nanoparticle labeled IgG were passed through the immunoaffinity column mounted in a flow system and competed for the surface-confined anti-IgG antibody. CL emission was generated from the reaction between luminol and hydrogen peroxide in the presence of Au (III), generated from chemically oxidative dissolution of gold nanoparticle by an injection of 0.10 mol L−1 HCl–0.10 mol L−1 NaCl solution containing 0.10 mmol L−1 Br2. The concentration of analyte IgG was inversely related to the amount of bound gold nanoparticle labeled IgG and the CL intensity was linear with the concentration of analyte IgG from 1.0 ng mL−1 to 40 ng mL−1 with a detection limit of 5.2 × 10−10 g mL−1. The whole assay time including the injections and washing steps was only 30 min for one sample, which was competitive with CL immunoassays based on a gold nanoparticle label and magnetic separation. This work demonstrates that the CL immunoassay incorporation of nanoparticle label and flow injection is promising for clinical assay with sensitivity and high-speed.

Graphical abstractFigure optionsDownload full-size imageDownload as PowerPoint slideHighlights► A sensitive competitive flow injection chemiluminescence immunoassay for immunoglobulin G was developed using gold nanoparticle as CL label. ► The sensitivity of CL immunoassay was much improved because of the release of a large number of Au (III) from each gold nanoparticle used as label. A lower detection limit of 5.2 × 10−10 g mL−1 was obtained for IgG. ► The developed competitive CL-FIA immunoassay of IgG is easily performed with high rapidity because of employing flow injection system. The whole assay time including the injections and washing steps was only 30 min for one sample.