Renewable Amperometric Immunosensor for Schistosoma japonium Antibody Assay

Abstract

A renewable amperometric immunosensor has been proposed for the determination of Schistosoma japonium antibody (SjAb) in rabbit serum. A paraffin−graphite−Schistosoma japonium antigen (SjAg) biocomposite, which needs no additional curing, was directly used to construct the immunosensors. The analytical sample containing the desired SjAb was mixed with SjAb labeled with horseradish peroxidase (HRP) to form the incubation solution for the competitive binding assay. Amperometry was used to determine the amount of HRP fixed on the sensor surface, which was related to the content of desired SjAb. Assay conditions were optimized, including the selection of substrate, the loading of SjAg in the biocomposite, the amount of labeled SjAb in the incubation solution, the incubation time, and the temperature. Using o-aminophenol (o-AP) as a substrate, amperometric detection at −200 mV (vs SCE) resulted in a pseudolinear detection range of about 0.36 to 14 μg/mL, with a detection limit of 0.36 μg/mL. Rabbit serum samples with varying infection degrees were analyzed, and the results demonstrated that the concentration that is detectable in this system meets the demands of clinical analyses. A new surface on the immunosensor for use in another competitive assay can be obtained by removing the original one and polishing the surface.