Antimicrobial use in livestock has emerged as a pressing global issue because of the rise of antimicrobial-resistant bacteria. Regulatory authorities across the globe have taken steps to discourage the misuse of these antibiotics by banning or limiting the use of medically important antibiotics in food animals. However, to ensure that food animals are not being administered antibiotics inappropriately, there is a need for a reliable, raid-response biosensor that can detect the presence of these antibiotic residuals in meat products. We have developed an affinity-based electrochemical biosensor for the label-free detection of ceftiofur residues in meat samples. The sensor uses a self-assembled immunoassay to target the ceftiofur biomarker by employing electrochemical impedance spectroscopy to probe the interfacial capacitive changes as ceftiofur binds to the sensor surface. We have demonstrated a platform that can detect ceftiofur within 15 min of introducing the sample at concentrations down to 0.01 ng/mL in 1× phosphate-buffered saline and 10 ng/mL in 220 mg ground turkey meat samples.