Thick-film electrochemical immunosensor based on stripping potentiometric detection of a metal ion label

A disposable electrochemical immunosensor based on potentiometric stripping analysis (PSA) of a metal tracer and using an entirely on-chip assay format is demonstrated. Challenges associated with the adaptation of earlier stripping voltammetric immunoassays to an on-chip operation, and with meeting the demands of decentralized testing, have been addressed. These include the surface immobilization of the antibody, the replacement of mercury drop electrodes, elimination of the separation and oxygen-removal steps, and the use of quiescent 30-microL sample droplets. Human serum albumin (HSA) and anti-HSA antibody were used as a model system, while bismuth ion served as the metal label. The anti-HSA was immobilized onto the surface of a thick-film electrode, followed by a competition between the Bi-labeled analyte-tracer and the analyte (HSA) for the antibody binding sites. Upon removal of the unbound tracer, Bi3+ was released and detected by PSA. The dynamic concentration range for HSA (0.3-30 micrograms/mL) and the detection limit (0.2 microgram/mL, i.e., 90 fmol in the 30-microL sample) indicate that the greatly simplified protocol does not compromise the performance characteristics of stripping immunoassays. Consequently, this on-chip operation offers great promise for decentralized (clinical and environmental) applications.