Serum bilirubin is an important biomarker for the diagnosis of various types of liver diseases and blood disorders. A polydopamine/gold nanoclusters composite film was fabricated for the fluorescent sensing of free bilirubin. Bovine serum albumin (BSA)-stabilized gold nanoclusters (AuNCs) were used as probes for biorecognition. The polydopamine film was utilized as an adhesion layer for immobilization of AuNCs. When the composite film was exposed to free bilirubin, due to the complex that was formed between BSA and free bilirubin, the fluorescence intensity of the composite film was gradually weakened as the bilirubin concentration increased. The fluorescence quenching ratio (F0/F) was linearly proportional to free bilirubin over the concentration range of 0.8~50 μmol/L with a limit of detection of 0.61 ± 0.12 μmol/L (S/N = 3). The response was quick, the film was recyclable, and common ingredients in human serum did not interfere with the detection of free bilirubin.