Enzymatic biosensing by covalent conjugation of enzymes to 3D-networks of graphene nanosheets on arrays of vertically aligned gold nanorods: Application to voltammetric glucose sensing.

The authors demonstrate efficient direct electron transfer from the enzyme glucose oxidase to vertically aligned gold nanorods with a diameter of ~160 nm and a length of ~2 μm that are covalently linkage to a 3-dimensional network of reduced graphene oxide nanosheets. The assembly can be prepared by a 2-step electrochemical procedure. This hybrid structure holds the enzyme in a favorable position while retaining its functionality that ultimately provides enhanced performance for enzymatic sensing of glucose without utilizing mediators. The nanorod assembly was applied to the voltammetric detection of glucose. Figures of merit include an electrochemical sensitivity of 12 μA·mM-1·cm-2 (obtained from cathodic peak current at a voltage of -0.45 V vs. Ag/AgCl), a 3 μM detection limit (at signal/noise = 3), and a wide linear range (0.01-7 mM). The hybrid nanostructure has a heterogeneous electron transfer rate constant (ks) of 2.9 s-1. The high electrochemical activity is attributed to the synergistic effect of a large active surface and an enhanced electron transfer efficiency due to covalent amide linkage. Graphical Abstract Schematic of the procedure utilized for the fabrication of an electrochemical biosensor based on gold nanorods (AuNRs) modified with a reduced graphene oxide (rGO)/glucose oxidase (GOx) conjugate. The enzyme electrode was employed to the determination of glucose by differential pulse voltammetry.