Bacterial biofilms are a leading cause of infection in health-care settings. Surface plasmon resonance (SPR) biosensors stand as valuable tools not only for the detection of biological entities and the characterisation of biomaterials but also as a suitable means to monitor bacterial film formation. This article reports on a proof-of-concept study for the use of an angular-based SPR biosensor for the monitoring of bacterial cell growth and biofilm formation and removal under the effect of different cleaning agents. The benefit of this custom-made SPR instrument is that it records simultaneously both the critical and resonant angles. This provides unique information on the growth of bacterial cells which is otherwise not obtainable with commonly used intensity-based SPR systems. The results clearly showed that a multilayer biofilm can be formed in 48 hours and the steps involved can be monitored in real-time with the SPR instrument through the measurement of the refractive index change and following the evolution in the shape of the SPR curve. The number, the depth and the sharpness of the reflection ripples varied as the film became thicker. Simulation results confirmed that the number of layers of bacteria affected the number of ripples at the critical angle. Real-time monitoring of the film breakdown with three cleaning agents indicated that bleach solution at 4.5% was the most effective in disrupting the biofilm from the gold sensor. Our overall findings suggest that the SPR biosensor with angular modulation presented in this article can perform real-time monitoring of biofilm formation and has the potential to be used as a platform to test the efficiency of disinfectants.