Adalimumab is a humanized IgG1 monoclonal antibody (Mab) used to treat rheumatoid arthritis, inflammatory bowel diseases, Crohn’s disease, cancer and skin diseases. This drug specifically targets tumor necrosis factor alpha (TNF-α) by blocking its interaction with cell surface TNF-α receptors p55 and p75. Adalimumab diminishes inflammation, immunological reactions and thus minimizes tissue damage in autoimmune and inflammatory disorders. Of note, development of appropriate positive and negative controls is crucial for assay validation, interpretation and distinguishing antibody positive from antibody negative samples. Therefore, this study aimed to develop a bridge ELISA method using rabbit anti-adalimumab antibody as a positive control to screen anti-drug antibodies (ADAs). The positive control antibody was prepared by depleting human IgG specific antibodies and characterized for its specific reactivity with adalimumab, and without cross reactivity with human IgG. The method was developed and validated for the drug tolerant level by spiking the various concentrations of adalimumab with constant amount of rabbit anti-adalimumab antibody. The assay cut-off was determined by analyzing the mean concentration value of negative control + (2 x standard deviation). Remarkably, the precise identification and surveillance of ADAs are essential for enhancing biologic treatments and controlling immunogenicity. Moreover, the bridge ELISA has proven to be efficient among various approaches, due to its robustness, high sensitivity, and tolerance for variations in drug concentration. Of note, the newly designed bridge ELISA for anti-adalimumab antibodies exhibits superior performance, capable of detecting low ADA levels (0.0244 µg/mL) and conforming to rigorous EMA validation standards.