Abstract: An efficient approach to obtaining molecularly imprinted polymer (MIP) film for detection of 17β-Estradiol (E2) with water-compatible property, using a reversible addition-fragmentation chain transfer (RAFT) by photo-initiated is described. 2-methyl-2-[(dodecylsulfanylthiocarbonyl)sulfanyl]propanoic acid (DDMAT) as the sole control agent in the absence of additional photoinitiators or catalysts was utilized for the synthesis of MIP film. The MIP film was characterized by contact angle measurements, infrared spectroscopy (IR) and scanning electron microscope (SEM). Analysis of surface plasmon resonance (SPR) spectroscopy showed that the imprinted film displayed good sensitivity and selectivity to E2 compared to other competitors and non-imprinted polymer (NIP) film. The response of the E2 ranged from 10-14 to 10-6 mol/L with a limit of detection (LOD) of 1.15 x 10-15 mol/L (S/N=3) in PBS buffer (pH 7.4). This sensor showed a good reusability and stability, and kept about 84.8% of its original response after 45 days of storage in dry and ambient conditions. Finally, the sensor was applied to determine the E2 concentration in tap water and underground water samples in the range from 10-13 mol/L to 10-7 mol/L and the LODs were obtained as 7.4 x 10-15 mol/L and 1.12 x 10-14 mol/L, respectively