Surface plasmon resonance (SPR) biosensors with unique features of label-free, real-time and noncontact detection have been successfully used for the quantitative measurement of dynamics of molecular interactions in analytical chemistry and clinical biochemical analysis. In this paper, we discussed recent advancements in SPR biosensors to overcome several limitations of traditional devices and systems. We presented four examples, i.e., spacer layer design, surface immobilization strategy, sample handling with lateral flow layer, and isothermal thermal amplification for HBV detection to demonstrate the many possible configurations of SPR devices and systems for practical applications.