Immobilization of uricase enzyme on self-assembled gold nanoparticles for application in uric acid biosensor.

An enzyme immobilization matrix is described by preparing a self-assembly of gold nanoparticles (GNPs) over a self-assembled monolayer (SAM) of 3-aminopropyltriethoxysilane (APTES) on an indium-tin-oxide (ITO) coated glass plate. The surface of the GNPs was modified with a mixed (1:9) SAM of 11-mercaptoundecanoic acid (MUA) and 3-mercapto-propionic acid (MPA). The enzyme, uricase was covalently immobilized to the carboxyl groups of the mixed SAM of MUA/MPA through carbodiimide coupling reaction. The whole assembly was constructed on 1 cm2 area of ITO-glass plate and was tested as an amperometric biosensor for the detection of uric acid in aqueous solution. The biosensor assembly was characterized by atomic force microscopy (AFM) and electrochemical techniques. The AFM of the enzyme biosensor assembly reveals an asymmetrical sharp regular island-like structure with an average roughness parameter value of 2.81 nm. Chronoamperometric response was measured as a function of uric acid concentration in aqueous solution (pH 7.4), which exhibits a linear response over a concentration range of 0.07 to 0.63 mM with a sensitivity of 19.27 microAmM(-1) and a response of 25 s with excellent reproducibility. These results are not influenced by the presence of interfering reagents such as ascorbic acid, urea and glucose. GNPs-biomolecule assemblies constructed using this method may facilitate development of new hybrid biosensing materials.