The formation of self-assembled monolayers (SAMs) is a normal method for the immobilization of biorecognition elements immobilized on SPR sensors. With this method, mercaptopropionic acid (MPA) with carboxylic and thiol group is the most commonly used. Dimercaptosuccinic acid (DMSA) having two carboxylics and two thiol groups is a classical antidote for heavy metal ions. In this paper, DMSA was first used to form SAMs to connect the antigen on the chip of a surface plasmon resonance (SPR) immunosensor for detection of salbutamol (SAL), and the results were compared with a traditional (MPA)-SPR sensor. Dihydrolipoic acid(DHLA)-SPR showed that the recognition efficiency of antigen and antibody of DMSA-SPR immunosensor was 170.1% at room temperature with the linear range of 5 – 150 ng/mL. The recovery rate of this sensor applied to SAL detection in pork reached 94.9 – 108.0% and the limit of quantification (LOQ) was 5 ng/mL. The results were in good correlation with the analysis results of ultra-high phase liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) analysis. This novel DMSA-SPR immunosensor provides insight into a new idea and method for improving the sensitivity of SPR immunosensors and can be widely used in the detection of other small molecules.