Quantum dots (QDs) luminescent films are extensively applied to optoelectronics and optical devices. However, QDs aggregation results in the quenching of their fluorescence property which limits their practical applications to a greater extent. In order to resolve this issue, 3-mercaptopropionic acid (3-MPA) functionalized Cadmium Tellurium (CdTe) QDs were stabilized by silk fibroin (SB) and co-assembled with layered doubled hydroxide (LDH) to form (QDs@SF/LDH)n ultrathin films (UTFs) via the layer-by-layer (LBL) technique. UV–Vis absorption and fluorescence spectroscopy showed a stepwise and normal growth of the films upon increasing the number of deposition cycles. XRD and AFM studies confirmed the formation of a periodic layered structure and regular surface morphology of the thin films. As compared to (CdTe QDs/LDH)nUTFs, the (CdTe QDs@SF/LDH)nUTFs displayed fluorescence enhancement and longer fluorescent lifetime, both in solid states and aqueous solutions. Furthermore compared with the solution state, the fluorescence enhancement of SF-RC and SF-β are, respectively, 7 times and 17 times in the (CdTe QDs@SF/LDH)n UTFs, indicating that the LDH nanosheets favor the fluorescence enhancement effect on the CdTe QDs@SF. The fabricated materials displayed fluorescence response to a biological molecule such as immune globulin, lgG. Thus, the (CdTe QDs@SF/LDH)n UTFs has a potential to be used as biosensor. Keywords: CdTe quantum dots, Silk fibroin, Layered doubled hydroxide, Co-assembly, Fluorescence enhancement