Abstract: A novel, sensitive anodic stripping volammetric immunoassay was developed based on silver-enhanced immuno-gold label. The immunoreaction was performed in a polystyrene microwell by using the sandwich format. Primary antibodies specific for C₃(complement Ⅲ) were adsorbed passively on the walls of a polystyrene microwell. The C₃ analyte was first captured by the primary antibody and then sandwiched by a secondary colloidal gold-labeled antibody. The addition of the silver enhancement solution results in the precipitation of a large amount of silver on colloidal gold labels due to the catalytic reduction which, after silver metal dissolution in an acidic solution, was determined by anodic stripping voltammetry(ASV) at a glassy-carbon electrode. The influence of some immunoassay conditions upon the anodic stripping peak current was examined and optimized. The anodic stripping peak current depended linearly on the logarithm of C₃ mass concentration over the range of 7.2 ng/mL to 7.33 μg/mL and a detection limit as low as 7ng/mL is achieved. The anodic stripping volammetric immunoassay was applied to the determination of C₃ concentration in human serum with satisfactory results.

Key words: Metalloimmunoassay, Anodic stripping voltammetry, Silver enhancement solution, Immuno-gold label