Numerous studies have been reported to improve the selectivity of uric acid (UA) by eliminating the interference from other electroactive species that coexist in biological fluids. However, two main challenges associated with the nonenzymatic electrochemical detection of UA need to be overcome to achieve practical applications in biological samples. Those are the chemical fouling of electrodes caused by the oxidation product of UA and biofouling due to the non-specific absorption of biological macromolecules. It was found that the residual oxo-functional groups and defects on graphene played a crucial part in both electrocatalysis and anti-biofouling. Here, graphene oxide (GO) was tuned by electro-oxidation and electro-reduction and was investigated in antifouling and electrocatalytic performances for the electrochemical sensing of UA by using pristine GO, BSA bound GO, electro-reduction-treated GO and electro-oxidation-treated GO. The electro-oxidation-treated GO was explored in electrochemical sensing for the first time and exhibited the highest sensitivity and low fouling properties. Holey GO might be formed on the electrode surface by the electrochemical oxidation method in a mild and green solution without the use of an acid. The different electrode interfaces as well as the interaction with BSA were investigated by Raman spectroscopy, X-ray photoelectron spectroscopy, contact angle measurements, scanning electron microscopy, electrochemistry, and electrochemical impedance spectroscopy.