Potential control of horseradish peroxidase immobilization on gold electrode

A new approach based on potential control was firstly used for the immobilization of horseradish peroxidase (HRP) as the model protein. The self-assembly monolayer (SAM) was prepared with 2-aminoethanethiol (AET) on the gold electrode. The charge on HRP was adjusted by means of the acidity of the phosphate buffer solution (PBS) for dissolving the HRP. The influence of electric potential on HRP immobilization was investigated by means of colorimetric immunoassay of enzyme-substrate interaction (CIESI) using an automatic plate reader. The HRP modified electrodes were characterized with X-ray photoelectron spectroscopy (XPS) as well as atomic force microscope (AFM) on template-stripped gold surface. The potential for maximum immobilization of HRP was near the zero charge potential. The result indicates that controlled potential can affect the course of HRP immobilization without the loss of enzymic activity. It is of great significance for the control of biomolecular self-assembly and the intrinsic electric device.