Renewable amperometric immunosensor for complement 3 assay based on the sol–gel technique

Abstract

A renewable amperometric immunosensor based on the sol–gel technique has been constructed by dispersing graphite, complement 3 (C3) antiserum, and sol–gel at low temperature. The prepared immunosensor is rigid, porous, and has a renewable external surface. A competitive binding assay has been used to determine C3 in human serum with the aid of C3 labeled with horseradish peroxidase. The enzyme-labeled antigen can readily diffuse toward the encapsulated antibody, which retains its binding properties. The experimental conditions for the assay with the biocomposite, including the loading of C3 antiserum in the biocomposite, the amount of labeled C3 in incubation solution, incubation time, and temperature, have been optimized. Using C3 labeled with horseradish peroxidase, and o-AP as the substrate, amperometric detection at –150 mV (relative to the SCE) results in a linear detection range of 1.17–35.1 μg mL^–1, with a detection limit of 0.56 μg mL^–1. Serum samples have been assayed and the results demonstrate the feasibility of the proposed immunosensor for clinical analysis. The surface of the immunosensor can be renewed simply by polishing to obtain a fresh immunocomposite ready to use in a new competitive assay.