Surface plasmon resonance (SPR) is a real-time, label-free, and high-sensitive detection technology. SPR has been widely used in many applications such as biomolecular interaction analysis, environmental monitoring, and medical diagnostics. However, conventional SPR sensor systems usually require expensive equipment and complicated optics. In this paper, we have demonstrated a rapid prototyping of angle-scanning SPR for bioanalytical investigation. Rapid prototyping was attained by utilizing the FDM (fused deposition modeling) based 3D (three-dimensional) printing technology. Two rotating platforms were employed to drive the laser source and photodiode, respectively. A temperature regulation unit was incorporated to maintain the system temperature in order to reduce the temperature effect. The proposed SPR rapid prototyping yielded a refractive index resolution of 6.4×10 −6 RIU (refractive index unit), and the biotin-avidin system validated the kinetics parameters measurement capability. The obtained results indicated that the FDM 3D printing has great potential for developing rapid-prototyping SPR system.