Electrochemical Immunosensor for Cholera Toxin Using Liposomes and Poly(3,4-ethylenedioxythiophene)-Coated Carbon Nanotubes

Abstract

A sensitive method for the detection of cholera toxin (CT) using an electrochemical immunosensor with liposomic magnification followed by adsorptive square-wave stripping voltammetry is described. Potassium ferrocyanide-encapsulated and ganglioside (GM1)-functionalized liposomes act as highly specific recognition labels for the amplified detection of cholera toxin. The sensing interface consists of monoclonal antibody against the B subunit of CT that is linked to poly(3,4-ethylenedioxythiophene) coated on Nafion-supported multiwalled carbon nanotube caste film on a glassy carbon electrode. The CT is detected by a "sandwich-type" assay on the electronic transducers, where the toxin is first bound to the anti-CT antibody and then to the GM1-functionalized liposome. The potassium ferrocyanide molecules are released from the bounded liposomes on the electrode by lyses with methanolic solution of Triton X-100. The released electroactive marker is measured by adsorptive square-wave stripping voltammetry. The sandwich assay provides the amplification route for the detection of the CT present in ultratrace levels. The calibration curve for CT had a linear range of 10(-14)-10(-7)g mL(-1). The detection limit of this immunosensor was 10(-16) g of cholera toxin (equivalent to 100 microL of 10(-15) g mL(-1)).