Target-induced synergetic modulation of electrochemical tag concentration and electrode surface passivation for one-step sampling filtration-free detection of acid phosphatase activity

Abstract

As a biomarker of several diseases, the activity of acid phosphatase (ACP) is generally used to assistantly diagnose these diseases. Thus, developing reliable ACP activity analytical methods becomes quite significant. Herein, we recommend a one-step sampling filtration-free electrochemical method for ACP activity determination based on the target-induced synergetic modulation of tag concentration and surface passivation. Mn3O4 microspheres with favorable oxidase-mimicking activity are synthesized to catalyze the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) to its product TMBox, resulting in a remarkable re-reduction signal of TMBox to TMB recorded by an integrated electrochemical system consisting of screen-printed electrode (SPE) and 3D-printed holder. When hexametaphosphate ions (HMPi) with rich negative charges are employed to interact positively charged TMBox, the formed flocculent precipitate TMBox-HMPi automatically sedimentates onto SPE surface, and both the decreased concentration of free TMBox in solution and the increased electrode surface passivation triggered by TMBox-HMPi sedimentation synergistically reduce the re-reduction signal of TMBox. When ACP is present, it hydrolyzes the HMPi substrate, greatly relieving the formation of the TMBox-HMPi precipitate and its sedimentation onto SPE surface. As a result, the electrochemical re-reduction signal of TMBox becomes remarkable again. With the strategy of using one stimulus to generate two-fold signal change, highly sensitive ACP activity detection was realized, with a wide linear range from 0.05 to 50 U/L and a limit of detection down to 0.024 U/L. Reliable monitoring of ACP activity in clinical serum was also demonstrated.