Breast cancer is the most common cause of cancer death in women; therefore, its early detection and treatment are crucial. To achieve this goal, we designed an optical sensor based on direct interaction of trastuzumab [Herceptin (HER)], a monoclonal antibody used to treat HER2-positive breast cancer, with plasmonic nanoparticles. Surface-modified gold nanoparticles (AuNPs) have gained considerable attention in biosensing techniques over the last years, which actuated these nanoparticles to the heart of various biosensing notions. We have exploited the localized surface plasmon resonance (LSPR) of gold nanoparticles to determine HER in human serum. AuNPs were decorated with negatively charged citrate ions, yielding enhanced direct-surface interaction with HER antibodies. The AuNPs are mixed with silver nanoparticles (AgNPs) in an optimized ratio to increase selectivity and sensitivity further. AuNPs detect the HER antibodies using LSPR, whereas AgNPs help monitor interferences' effect on the sensing media. The three effective factors in HER sensing, including the nanoparticle ratio, temperature, and pH were optimized via response surface methodology (RSM) based on the central composite design (CCD). The sensor's response toward HER was achieved in the linear range of 0.5 × 10-7 to 40 × 10-7 M with the detection limit of 3.7 × 10-9 M and relative standard deviation (RSD) less than 5%. The selectivity of the LSPR sensor was assessed by monitoring its response toward HER in the presence of other biological molecules with similar physicochemical properties. Rapid response time (less than 1 min), selectivity, and the simplicity of the developed LSPR-based sensor are the key advantages of the developed sensor.