Amperometric cholesterol biosensors based on hybrid organic–inorganic Langmuir–Blodgett films

Preparation and characterization of a thin-film cholesterol biosensor employing an organic–inorganic hybrid system of cholesterol oxidase (ChOx) and Prussian blue (PB) is described. ChOx was immobilized in Langmuir–Blodgett (LB) films consisting of positively charged octadecyltrimethylammonium (ODTA) and nano-sized PB clusters. Immobilization was performed by simple immersion of ODTA/PB LB films into an aqueous solution of ChOx. Subsequent ChOx absorption into LB films was confirmed by infrared reflection–absorption spectroscopy. Obtained ODTA/PB/ChOx LB films clearly exhibited a response current to cholesterol under an applied potential of 0.0 V (vs. Ag/AgCl). The linearity of current density versus cholesterol concentration was confirmed for the range 0.2–1.2 mmol/L. The present study indicates that simple immersion of ODTA/PB LB films into an enzyme solution is a promising method to produce many types of thin-film biosensors comprising a hybrid system of an oxidative enzyme and PB nano-clusters that work at a very low potential range.