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In this paper, we describe a chitosan-coated multi-wall carbon nanotube film which was used as a highly selective and sensitive biosensor for urea determination. Urease immobilization on multi-wall carbon nanotube films was achieved by crosslinking chitosan with glutaraldehyde (GA). The immobilized urease showed increased urea enzymatic activity of approximately 20-fold compared with the free enzyme in solution. The glutaraldehyde-induced crosslinking of chitosan significantly increases the film thickness and decreases its solubility in aqueous medium. Thus, a chitosan-coated multi-wall carbon nanotube film, which was thicker and more hydrophilic than a chitosan film, showed improved enzyme immobilization. The use of this kind of chitosan-coated multi-wall carbon nanotube film as a urea biosensor provided a sensitive (30 nM) and a selective response (approximately 20-fold higher sensitivity than the response to pyruvate). The response time and the reaction linear range were also significantly improved. The limit of detection of the urea biosensor was 0.11 microM. The urea biosensor displayed a fast response to urea concentration changes. The apparent Michaelis-Menten constant of the immobilized urease in the biosensor was 25 mM.Q:
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