A new patterning method is proposed for selective deposition of organic/organic and organic/inorganic multilayers by using an enzymatic surface. It was observed that the method offers an electrostatic charge reversal through pH control. It was observed during the study that a high-performance electrochemical sensors with catalytic properties can be prepared with this method. The study also found that the selective growth of multilayers by electrostatic repulsion can be achieved on enzyme-coated substrates. The direct contact between the probe molecules and the catalytic components can cause of an increase of electrochemical activity. The study used an electrophoretic light scattering spectrophotometer to determine the CAT after the adsorption and atomic force microscopy (AFM) to investigate multilayers.