Automated flow-through amperometric immunosensor for highly sensitive and on-line detection of okadaic acid in mussel sample

Abstract

An electrochemical immunosensor for okadaic acid (OA) detection has been developed, and used in an indirect competitive immunoassay format under automated flow conditions. The biosensor was fabricated by injecting OA modified magnetic beads onto screen printed carbon electrode (SPCE) in the flow system. The OA present in the sample competed with the immobilized OA to bind with anti-okadaic acid monoclonal antibody (anti-OA-MAb). The secondary alkaline phosphatase labeled antibody was used to perform electrochemical detection. The current response obtained from the labeled alkaline phosphatase to 1-naphthyl phosphate decreased proportionally to the concentration of free OA in the sample. The calculated limit of detection (LOD) was 0.15 μg/L with a linear range of 0.19-25 μg/L. The good recoveries percentages validated the immunosensor application for real mussel samples. The developed system automatically controlled the incubation, washing and current measurement steps, showing its potential use for OA determination in field analysis.