Multiplexed Ultrasensitive Determination of Adrenocorticotropin and Cortisol Hormones at a Dual Electrochemical Immunosensor

A novel dual electrochemical immunosensor for the multiplexed determination of adrenocorticotropin (ACTH) and cortisol is reported. Aminophenylboronic acid-modified dual screen-printed carbon electrodes were prepared on which the corresponding ACTH and cortisol antibodies were immobilized. Competitive immunoassays involved biotinylated ACTH and alkaline phosphatase labelled streptavidin, or alkaline phosphatase labelled cortisol. Differential pulse voltammetry upon 1-naphtyl phosphate addition was employed to monitor the affinity reactions. The ranges of linearity were 5.0X10-5-0.1 and 0.1-500 ng/mL for ACTH and cortisol. The usefulness of the dual immunosensor was demonstrated by analyzing certified human serum samples with good recoveries.