Biomolecular urease thin films grown by laser techniques for blood diagnostic applications

Matrix assisted pulsed laser evaporation (MAPLE) was used for growing urease thin films designed for bio-sensor applications in clinical diagnostics. The targets exposed to laser radiation were made from a frozen composite manufactured by dissolving biomaterials in distilled water. We used a UV KrF* (λ = 248 nm, τFWHM ≅ 30 ns, ν = 10 Hz) excimer source for multipulse laser irradiation of the frozen targets cooled with Peltier elements. The laser source was operated at an incident fluence of 0.4 J/cm2. Urease activity and kinetics were assayed by the Worthington method that monitors urea hydrolysis by coupling ammonia production to a glutamate dehydrogenase reaction. A decrease in absorbance was measured at 340 nm and correlated with the enzymatic activity of urease. We show that the urease films obtained by MAPLE techniques remain active up to three months after deposition.