The fate and transport of nanoparticles (NPs) in streams is critical for understanding their overall environmental impact. Using a unique field-scale stream at the Notre Dame-Linked Experimental Ecosystem Facility, we investigated the impact of biofilms and the presence of dissolved organic matter (DOM) on the transport of titanium dioxide (TiO2) NPs. Experimental breakthrough curves were analyzed using temporal moments and fit using a mobile-immobile model. The presence of biofilms in the stream severely reduced the transport of the TiO2 NPs, but this was mitigated by the presence of DOM. Under minimal biofilm conditions, the presence of DOM increased the mass recovery of TiO2 from 4.2% to 32% for samples taken 50 m downstream. For thriving biofilm conditions only 0.5% of the TiO2 mass was recovered (50 m), but the presence of DOM improved the mass recovery TiO2 to 36%. The model was suitable for predicting early, peak, tail, and truncation time portions of the breakthrough curves, which attests to its ability to capture a range of processes in the mobile and immobile domains of the stream. The model outcomes supported the hypothesis that DOM changed the interaction of NP-biofilm from an irreversible to a reversible process. Collectively, these outcomes stress the importance of considering biogeological complexity when predicting the transport of NPs in streams.