In this study, we developed a reflective localized surface plasmon resonance (LSPR) optical fiber sensor, based on silver nanoparticles (Ag NPs). To enhance the sensitivity of the LSPR optical sensor, two key parameters were optimized, the length of the sensing area and the coating time of the Ag NPs. A sensing length of 1.5 cm and a 1-h coating time proved to be suitable conditions to produce highly sensitive sensors for biosensing. The optimized sensor has a high refractive index sensitivity of 387 nm/RIU, which is much higher than that of other reported individual silver nanoparticles in solutions. Moreover, the sensor was further modified with antigen to act as a biosensor. Distinctive wavelength shifts were found after each surface modification step. In addition, the reflective LSPR optical fiber sensor has high reproducibility and stability.