Immunoassay of C-reactive protein by hot electron-induced electrochemiluminescence at polystyrene-carbon black composite electrodes

Aromatic Tb(III) chelates can be detected down to subnanomolar concentrations at polystyrene-carbon black composite electrodes on the basis of hot electron-induced electrochemiluminescence, and used as electrochemiluminescent labels in bioaffinity assays. The excitation mechanism is based on chemiluminescent reactions that are initiated by the field emission/tunnel emission of hot electrons from the composite electrode. The composition and the properties of the novel composite electrodes were studied in detail. Hot electron-induced electrochemiluminescence intensities obtainable with the present composite electrodes are comparable to the previously used metal/insulator-type electrodes without exhibiting considerable long-lived solid state electroluminescence background emission, unlike e.g. oxide-coated silicon and aluminum electrodes. C-reactive protein was finally determined in a heterogeneous sandwich-assay by using the present composite electrodes as a solid support material for the capture antibodies. The detection antibody was labeled with a commercially available Tb(II) chelate. Calibration curve for the determination of C-reactive protein was linear over two orders of magnitude and the detection limit was well below the clinical reference value, approx. 1 μg l⁻¹. Disposable polystyrene-carbon black composite electrodes are superior alternatives for e.g. silicon-based electrodes due to the low cost, easier manufacturing process and a very good performance e.g. in immunoassays.