A novel biosensor (immuno-rotary biosensor) was developed by conjugating deoxynivalenol (DON) monoclonal antibodies with the “rotator” ε-subunit of F 0 F 1 –ATPase within chromatophores with an ε-subunit monoclonal antibody–biotin–avidin–biotin linker to capture DON residues. The conjugation conditions were then optimized. The capture of DON was based on the antibody–antigen reaction and it is indicated by the change in ATP synthetic activity of F 0 F 1 –ATPase, which is measured via chemiluminescence using the luciferin–luciferase system with a computerized microplate luminometer analyzer. 10 −7 mg/ml of DON can be detected. The whole detection process requires only about 20min. This method has promising applications in the detection of small molecular compounds because of its rapidity, simplicity, and sensitivity.