A prismatic excitation-based affinity biosensor consisting of the prism (BK7), WS2/graphene hybrid nanosheets, and silver (Ag) as the active metal for the surface plasmon resonance is proposed in this present research. The introduction of the transition metal WS2/graphene layer protected the silver substrate and enhanced the adsorption of biomolecules, which facilitated the quality and performance of detection. Here, we improved the detection structure by focusing on the metallic materials, graphene and WS2 film layers, and the thickness of the measured medium on the sensing effect. The results show that the silver film had a more desirable resonance effect, and the design of the symmetric detection structure produced a double resonance peak, and it provides a reference for distributed sensing. Changing the thickness of the detection medium can dynamically adjust the wave vector matching conditions, which gives the sensor a certain tunability. In the bilayer WS2 and monolayer graphene (W = 2, G = 1) configuration, the sensitivity was up to 224 deg/RIU with a quality factor of 96.97 RIU−1, which has potential for clinical analytic and biochemical detecting applications.