A Colorimetric Enzyme-Linked Immunosorbent Assay with CuO Nanoparticles as Signal Labels Based on the Growth of Gold Nanoparticles In Situ

A colorimetric immunoassay has been reported for prostate-specific antigen (PSA) detection with CuO nanoparticles (CuO NPs) as signal labels. The method is based on Cu²⁺-catalyzed oxidation of ascorbic acid (AA) by O₂ to depress the formation of colored gold nanoparticles (AuNPs). Specifically, HAuCl₄ can be reduced by AA to produce AuNPs in situ. In the presence of target, CuO NPs-labeled antibodies were captured via the sandwich-type immunoreaction. After dissolving CuO nanoparticles with acid, the released Cu²⁺ catalyzed the oxidation of AA by O₂, thus depressing the generation of AuNPs. To demonstrate the accuracy of the colorimetric assay, the released Cu²⁺ was further determined by a fluorescence probe. The colorimetric immunoassay shows a linear relationship for PSA detection in the range of 0.1~10 ng/mL. The detection limit of 0.05 ng/mL is comparable to that obtained by other CuO NPs-based methods. The high throughput, simplicity, and sensitivity of the proposed colorimetric immunoassay exhibited good applicability for assays of serum samples.