We report the enzymatic cleavage of fluorescein diphosphate (FDP) by alkaline phosphatase (AP) confined in the tip of a nanopipet with a volume of approximately 100 attoliters. The amount of enzyme that reversibly adsorbs in the pipet tip was shown to be proportional to the enzyme concentration in the bath. Increasing the voltage applied to the pipet, up to 1.5 V, linearly controls the substrate turnover rate by increasing the flow of substrate out of the pipet. This work opens up possibilities of highly miniaturized and sensitive enzyme assays with high spatial resolution.
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Bruckbauer et al. (2004) studied this question.
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