Biosensors that can automatically and continuously track fluctuations in biomarker levels over time are essential for real-time sensing in biomedical and environmental applications. Although many electrochemical sensors have been developed to quickly and sensitively monitor biomarkers, their sensing stability in complex biofluids is disturbed by unavoidable nonspecific adhesion of proteins or bacteria. Recently, various substrate surface modification techniques have been developed to resist biofouling, yet functionalization of electrodes in sensors to be anti-biofouling is rarely achieved. Here, we report an integrated three-electrode system (ITES) modified with a "liquid-like" polydimethylsiloxane (PDMS) brush that can continuously and stably monitor reactive oxygen species (ROS) in complex fluids. Based on the slippery "liquid-like" coating, the modified ITES surface could prevent the adhesion of various liquids as well as the adhesion of proteins and bacteria. The "liquid-like" coating does not significantly affect the sensitivity of the electrode in detecting ROS, while the sensing performance could remain stable and free of bacterial attack even after 3 days of incubation with bacteria. In addition, the PDMS brush-modified ITES (PMITES) could continuously record ROS levels in bacterial-rich fluids with excellent stability over 24 h due to the reduced bacterial contamination on the electrode surface. This technique offers new opportunities for continuous and real-time monitoring of biomarkers that will facilitate the development of advanced sensors for biomedical and environmental applications.