Indoor allergen assessment quantified by a thin-layer electrochemical cell and magnetic beads

We report the electrochemical determination of mite allergen in real house dust by using a thin layer electrochemical flow cell and magnetic beads. Der p1, which is an allergen from Dermatophagoides pteronyssinus, was immunochemically sandwiched between two dispersed monoclonal antibodies; one was modified on the surface of magnetic beads and the other was modified with alkaline phosphatase. After washing the beads, a small volume of p-aminophenol phosphate (p-APP) was added to produce p-aminophenol (p-AP). And then the p-AP concentration was measured electrochemically with a homemade electrochemical cell. The Der p1 assay was completed within 30 min and a low detection limit of 0.3 ng/mL was achieved. This is because the diffusion distance of Der p1 and the detection antibody was reduced to 22.3 μm by using dispersed magnetic beads. Only 10 min was required to complete the entire immunoreaction, and 54% of the Der p1 was confirmed to have immunoreacted in only 1 min of mixing. Furthermore, the p-APP volume could be reduced using the thin-layer electrochemical flow cell. This is advantageous in terms of concentrating p-AP, and provides a high signal-to-noise ratio measurement in a short time. We achieved a high correlation (r=0.967, p<0.001) between our assay and a conventional enzyme-linked immunosorbent assay (ELISA) for real house dust measurements.