Two-dimensional (2D) transition-metal dichalcogenides have become promising candidates for surface-enhanced Raman spectroscopy (SERS), but currently very few examples of detection of relevant molecules are available. Herein, we show the detection of the lipophilic disease marker β-sitosterol on fewlayered MoTe 2 films. The chemical vapor deposition (CVD)grown films are capable of nanomolar detection, exceeding the performance of alternative noble-metal surfaces. We confirm that the enhancement occurs through the chemical enhancement (CE) mechanism via formation of a surface−analyte complex, which leads to an enhancement factor of ≈10 4 , as confirmed by Fourier transform infrared (FTIR), UV−vis, and cyclic voltammetry (CV) analyses and density functional theory (DFT) calculations. Low values of signal deviation over a seven-layered MoTe 2 film confirms the homogeneity and reproducibility of the results in comparison to noble-metal substrate analogues. Furthermore, β-sitosterol detection within cell culture media, a minimal loss of signal over 50 days, and the opportunity for sensor regeneration suggest that MoTe 2 can become a promising new SERS platform for biosensing.