Acetylcholine biosensor based on the electrochemical functionalization of graphene field-effect transistors

We present a new strategy of Acetylcholinesterease (AchE) immobilization on graphene field-effect transistors (gFETs) for building up Acetylcholine sensors. This method is based on the electrosynthesis of an amino moiety-bearing polymer layer on the graphene channel. The film of the copolymer poly(3-amino-benzylamine-co-aniline) (PABA) does not only provide the suitable electrostatic charge and non-denaturing environment for enzyme immobilization, but it also improves the pH sensitivity of the gFETs (from 40.8 to 56.3 μA/pH unit), probably due to its wider effective pKa distribution. The local pH changes caused by the enzyme-catalyzed hydrolysis produce a shift in the Dirac point of the gFETs to more negative values, which are evidenced as differences in the gFET conductivity and thereby constituted the signal transduction mechanism of the modified transistors. In this way, the constructed biosensors showed a LOD of 2.3 μM and were able to monitor Ach in the range from 5 to 1000 μM in a flow configuration. Moreover, they showed a sensitivity of −26.6 ± 0.7 μA/Ach decade and also exhibited a very low RSD of 2.6%, revealing good device-to-device reproducibility. The biosensors revealed an excellent selectivity to interferences known to be present in the extracellular milieu, and the response to Ach was recovered by 97.5% after the whole set of interferences injected. Finally, the biosensors showed a fast response time, with an average value of 130 s and a good long-term response.