An electrochemical biosensor integrating immunoassay and enzyme activity analysis for accurate detection of active human apurinic/apyrimidinic endonuclease 1

Abstract

A novel electrochemical biosensing method that can take into account both immunoassay and enzyme activity analysis was reported in this work for determination of the enzymatically active human apurinic/apyrimidinic endonuclease 1 (APE1). The basic principle is to design and construct a DNA catalytic hairpin assembly (CHA) triggered by APE1 catalysis in enzyme activity analysis, and the assembled DNAs are labeled with electrochemically active CdS and PbS quantum dots to output electrochemical signals. In this system, the signal generation needs to satisfy both the conditions of immunological recognition and enzymatic catalysis, providing a basis for accurate analysis of active APE1. Results show that this method can reflect the regulation of the enzyme activity and can also distinguish APE1 from its isozymes with the same enzyme activity. The concept and successful implementation of this integrated system will contribute to the research and application of APE1 in biomedicine, and provide a reference for the accurate analysis of other enzymes.

Keywords: Catalytic hairpin assembly; Electrochemical detection; Enzyme activity analysis; Human apurinic/apyrimidinic endonuclease 1; Immunoassay.