We report that the end facet of an optical fiber can be coated with polyelectrolyte multilayers (PEM) of polycation (diallyldimethyl ammonium chloride) and polyanion (styrenesulfonate sodium salt) (PDDA+PSS)n (n is the number of bilayers), which functions effectively as a Fresnel-reflection based biosensor. The experimental setup includes a broadband light source, a 3dB coupler, and an optical spectrum analyzer. Biotin and streptavidin are deposited onto the multilayers-coated end facet sequentially. The light intensity change due to variation of external refractive index is monitored. When the concentrations of streptavidin changes from 0.1mg/ml to 1mg/ml, a linear relationship between the concentration of streptavidin and the reflected optical power at the wavelength of 1530nm is observed. The sensitivity increases from -1.6262×10-3 dB/ppm to -4.7852 ×10-3 dB/ppm, when the number of PEM increases from 1 to 2. Then we confirm the optimized numbers of bilayers of PEM are 5 through experiment. Selectivity and repeatability of our proposed optical fiber biosensor are verified. When bovine serum albumin (BSA) is added instead of streptavidin, the obtained spectra overlaps with that of biotin’s. The final end facet coated with PEM and biotin-streptavidin can be cleaned using microwave vibration or aqua regia. The microwave vibration method is utilized due to security concern. The optical spectra changes back to the initial one of the optical fiber in air. In conclusion, a Fresnel-reflection based optical fiber biosensor with good sensitivity, selectivity and repeatability is proposed. This biosensor has the advantages of simple structure, low cost and reliability.