In this research, a reproducible surface plasmon resonance (SPR) immunosensor based on inhibition format was fabricated for stable and sensitive detection of four sulfonamides (SAs) in animal-derived foods. The parameters in the fabrication and measurement process were optimized and discussed in details. The method using the proposed SPR immunosensor was validated and exhibited favorable performance for SAs residues detection in common animal-derived food products, as well as acceptable accuracy (pure milk 88.5-106.2%; egg 89.0-91.5%; chicken muscle 87.2-94.7%; beef 84.8-96.9%; fish 87.8-95.0%), precision (relative standard deviation (n = 3), pure milk 3.2-5.2%; egg 1.1-5.4%; chicken muscle 1.1-5.9%; beef 2.5-7.1%; fish 4.6-5.3%), and sensitivity (IC 15 , pure milk 59.6 ng mL − 1 , egg 56.1 ng mL − 1 , chicken muscle 66.9 ng mL −1 , beef 63.3 ng mL −1 , fish 62.7 ng mL −1 ). Each detection cycle could finish in less than 5 min, and each SPR chip could reuse 300 analysis cycles with response attenuation of 3.7%. These results have demonstrated that the proposed SPR immunosensor has offered an effective, accurate, sensitive, rapid, and low-cost methodology for SAs residue detection in food sample, and this method has great potential for the routine analysis of large numbers of samples on measuring different kinds of compounds.