Multilayers of hemoglobin were uniformly deposited on a pyrolytic graphite electrode surface through a layer-by-layer self-assembly approach. The well-known anionic polyelectrolyte, DNA, was used to attract cationic hemoglobin due to the strong electrostatic interaction between DNA and the protein. Interestingly, the electron transfer reactivity of hemoglobin was significantly improved in the film, which in turn facilitated the electrocatalytic reduction of nitric oxide.

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An unmediated NO biosensor was prepared by co-immobilizing haemoglobin with polyethyleneimine at a pyrolytic graphite electrode surface. Haemoglobin exhibits direct electrochemistry in the membrane, and this modified electrode shows high stability and catalytic activity towards the reduction of NO. A linear relationship between the catalytic current and the concentration of NO was observed in the concentration ranges 1.0 x 10(-7)-8.0 x 10(-6) M and 1.0 x 10(-5)-2.5 x 10(-5) M, with a detection limit of 4.0 x 10(-8) M.

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