The presence of antibiotics in milk is a significant concern for food safety and public health due to potential risks of antibiotic resistance and adverse effects on consumer health. This chapter reviews the application of immunosensors for the detection of antibiotics in milk and provides an overview of the current methodologies and advancements in this area. Immunosensors demonstrate high specificity and sensitivity through antigen–antibody interactions in the detection of trace amounts of antibiotics. This chapter focuses on the principles of immunosensor technology, covering the types of immunosensors in pharmaceutical analysis, which include enzyme-linked immunosorbent assays (ELISA), surface plasmon resonance, and electrochemical immunosensors. We focus next on the challenges for antibiotic detection in complex milk matrices, considering possible interference by other milk components and requirements of sample preparation techniques. An update on nanomaterials and signal amplification strategies is also presented regarding their contribution to the higher performance of immunosensors. Moreover, this chapter contains some information about the prospects this immunosensor technology is facing in the view of food safety and regulatory requirements in terms of integration into routine quality control processes. This chapter gives the general overview of antibiotic residues monitoring systems development and implementation for effective milk quality control process.