A portable multi-channel surface plasmon resonance (SPR) biosensor device using prism-based wavelength interrogation is presented. LEDs were adopted as a simple and inexpensive light source, providing a stable spectrum bandwidth for the SPR system. The parallel light was obtained by a collimated unit and illuminated on the sensing chip at a specific angle. A simple, compact and cost-effective spectrometer part constituted of a series of lenses and a prism was designed for the collection of reflected light. Using the multi-channel microfluidic chip as the sensing component, spectral images of multiple tests could be acquired simultaneously, improving the signal processing and detection throughput. Different concentrations of sodium chloride aqueous solution were used to calibrate the device. The linear detection range was 4.32 × 10 refractive index units (RIU) and the limit of detection was 6.38 × 10 RIU. Finally, the performance of the miniaturized SPR system was evaluated by the detection of immunoglobulin G (IgG).