Amperometric uric acid biosensors fabricated of various types of uricase enzymes

Preparation process of an enzyme-based bipotentiostatic amperometric uric acid sensor has been investigated. The suitability of three different Uricase (EC 1.7.3.3) enzymes (from porcine liver, Candida Utilis, Bacillus Fastidiosus) is described in this paper. The sensor fabricated of Uricase from Candida Utilis showed a linear response to uric acid in the 0-0.9 mM concentration range and the response current range was 0-3.3 μA. The sensor fabricated of Uricase from Bacillus Fastidiosus has been saturated at 0.72 mM and the response was not linear above 0.24 mM. The response current range was 0-0.9 μA. The sensor fabricated of Uricase from porcine liver has not given detectable electrical signal due to its very low specific activity. The substrate was prepared by screen printing on sintered alumina ceramic sheets using pastes of Au or Pd-Pt as working (W) and counter (C) and Pt-Ag as a reference (R) electrode. Galvanostatic electrocopolymerization of dodecyl sulfate doped poly-N-methyl-pyrrole (pNMPy) layer was used for enzyme immobilization. The layout of the sensor consists of four electrode surfaces (W₁, W₂, R, and C). By the bipotentiostatic technique, the two working electrodes (with and without the enzyme) are identically prepared and polarized, while the currents in the two circuits are measured simultaneously; thus, the current of the W₂-C circuit (I₂) can be substracted as a nonspecific background noise. The nonspecific oxidation of uric acid on the poly-N-methyl-pyrrole layer at 0.2 V has been demonstrated in oxygen bubbled buffer solution.