A triple read-out visible biosensing platform based on multifunctional nanozyme and bipolar electrode for multi-mode detection and imaging of CEA

Abstract

In this paper, a proposal of closed bipolar electrode (BPE) and nanozyme based multi-mode biosensing platform is first presented. As a novel integrated chip, multi-mode-BPE (MMBPE) combines enzyme-linked immunoassay (ELISA), electrochemiluminescence (ECL), ECL imaging and light emitting diode (LED) imaging, enabling highly sensitive triple read-out visible detection of cancer embryonic antigen (CEA). The ECL probe Ab₂@Au@Co₃O₄/CoFe₂O₄ hollow nanocubes (HNCs) with excellent peroxidase (POD) activity is introduced into the BPE cathode through immune adsorption. The Au@Co₃O₄/CoFe₂O₄ HNCs can increase the rate of hydrogen peroxide oxidation of TMB, thus promoting the reaction, and can be used for ELISA detection of CEA at different concentrations. The modification of the BPE sensing interface and reporting interface involved the introduction of the luminescent reagent Ru(bpy)₃²⁺ to the BPE anode. The decomposition rate of H₂O₂ increased under the catalytic action of Au@Co₃O₄/CoFe₂O₄ HNCs nanozyme, leading to an accelerated electron transfer rate in the MMBPE system and an enhanced ECL signal from Ru(bpy)₃²⁺. The LED imaging technology further provides a convenient and visible approach for CEA imaging in which no additional chemicals are needed. The integration of nanoenzymes as the catalytic core in MMBPE system provides impetus, while the combination of nanozymes with BPE expands the application of nanoenzymes in the field of biological analysis. The integration of intelligent chips with multiple modes of detection shows portable, miniaturized, and integrated excellent properties which meets the requirements of modern detection devices and thus offers a flexible approach for determination of nucleic acids, proteins, and cells.

Keywords: Bipolar electrode; ELISA; Electrochemiluminescence; Multi-mode biosensing; Nanozymes.