The development of graphene-based devices requires control over the functionalization and modification of the graphene surface. We studied the modification of the monolayer graphene surface with downstream oxygen plasma treatment to form graphene oxide (GO) for use in glucose biosensors. Amperometric and voltammetric methods were applied to GO for glucose oxidase (GOx) immobilization with a simultaneous reduction of GO to reduce graphene oxide. The fabricated glucose sensor shows a sensitivity of 0.118 μA mM −1 cm −2 and a detection limit of 0.0526 mM of glucose. This method provides fast and simultaneous immobilization of GOx and reduction of GO, and can be used in the fabrication of other electrochemical biosensors. Glucose oxidase has been recognized as an enzyme for monitoring glucose. In amperometric biosensors for glucose detection, GOx is immobilized on a nanomaterial surface and catalytically oxidizes glucose for high sensitivity and selectivity.9-11 However, enzyme immobilization is complicated by poor electrical communication of the active sites of the enzyme and the electrode surface, and by enzyme leaching. [12][13][14][15] Recently, graphene has been used to modify biosensors, offering advantages for enzyme immobilization on graphene surfaces. [16][17][18] Pristine graphene (PrG), a monolayer structure composed of one-atom thick two-dimensional honeycomb lattices of sp 2 carbon atoms, has extraordinary properties such as high intrinsic mobility and excellent thermal and electrical conductivity. [19][20][21][22] Although the bulk properties of graphene are very promising for a variety of applications, the development of graphene-based devices requires great control over the functionalization and modification of the graphene surface. Graphene oxide (GO) is one branch of functionalized graphene. GO is typically covalently functionalized with hydroxyl, epoxy, carbonyl, and carboxyl groups on its basal planes and at its edges, 23 resulting in a hybrid structure comprising a mixture of sp 2 and sp 3 hybridized carbon atoms. 24 GO sheets synthesized from graphite powders by the modified Hummers method 25 have been coated on carbon and Pt electrodes and used to immobilize GOx to develop glucose sensors. [26][27][28][29] On the other hand, monolayer graphene has been fabricated by chemical vapor deposition (CVD) on Cu and Ni substrates then transferred to other substrates for chemical and gas sensors. [30][31][32] Monolayer graphene has also been utilized for glucose sensor after immobilizing GOx to its surface through linker molecules. 33,34 With the aid of lithography and plasma etch techniques, monolayer-graphene-based devices can be patterned into arrays 32 and integrated with Si microelectronics technology. 35,36 In this work we synthesized GO from monolayer graphene by a downstream oxygen plasma. 37,38 The plasma-assisted technique has the advantages of short treatment time, an environmentally friendly manufacturing process, and the generation of numerous active species. 39 The immobilization of GOx en...