Electrochemical Immunosensor Based on PolythionineGold Nanoparticles for the Determination of Aflatoxin B1.docVIP

Sensors 2008, 8, 8262-8274; DOI: 10.3390/s8128262 OPEN ACCESS sensors ISSN 1424-8220 /journal/sensors Article Electrochemical Immunosensor Based on Polythionine/Gold Nanoparticles for the Determination of Aflatoxin B1 Joseph H.O. Owino, Omotayo A. Arotiba, Nicolette Hendricks, Everlyne A. Songa, Nazeem Jahed, Tesfaye T. Waryo, Rachel F. Ngece, Priscilla G .L. Baker and Emmanuel I. Iwuoha* SensorLab, Department of Chemistry, University of Western Cape, Private Bag X17, Bellville 7535, Cape Town, South Africa * Author to whom correspondence should be addressed; E-mail: eiwuoha@uwc.ac.za Received: 19 September 2008; in revised form: 1 December 2008 / Accepted: 2 December 2008 / Published: 15 December 2008 Abstract: An aflatoxin B1 (AFB1) electrochemical immunosensor was developed by the immobilisation of aflatoxin B1-bovine serum albumin (AFB1-BSA) conjugate on a polythionine (PTH)/gold nanoparticles (AuNP)-modified glassy carbon electrode (GCE). The surface of the AFB1-BSA conjugate was covered with horseradish peroxidase (HRP), in order to prevent non-specific binding of the immunosensors with ions in the test solution. The AFB1 immunosensor exhibited a quasi-reversible electrochemistry as indicated by a cyclic voltammetric (CV) peak separation (ΔEp) value of 62 mV. The experimental procedure for the detection of AFB1 involved the setting up of a competition between free AFB1 and the immobilised AFB1-BSA conjugate for the binding sites of free anti-aflatoxin B1 (anti-AFB1) antibody. The immunosensor’s differential pulse voltammetry (DPV) responses (peak currents) decreased as the concentration of free AFB1 increased within a dynamic linear range (DLR) of 0.6 - 2.4 ng/mL AFB1 and a limit of detection (LOD) of 0.07 ng/mL AFB1. This immunosensing procedure eliminates the need for enzyme-labeled secondary antibodies normally used in convent