Therapeutic proteins, such as infliximab, have revolutionized the treatment of many diseases during the last decade and more than 80 therapeutic proteins are currently approved for clinical use. However, all exogenous proteins have the potential to cause antibody formation. In order to ensure patient safety and the efficacy of therapeutic proteins, careful monitoring of the immunogenicity of therapeutic proteins is therefore necessary not only during preclinical trials, but also during the treatment of patients. Here, we report a clear-cut demonstration of the induction of anti-infliximab antibodies during the treatment of a patient with ankylosing spondylitis ( AS). Assessment of anti-infliximab antibodies in sera obtained at various time periods were performed using a highly specific double antigen assay system developed in our laboratory. Immunoreactivity was found to be solely specific for infliximab. Because all sera obtained from the patient were found to be negative for the presence of human anti-mouse antibody ( HAMA) and anti-human antibodies. The loss of effect of infliximab, as judged by observing the relapse of signs and symptoms of disease in the patient, seemed to be related with the appearance of antibodies. This study clearly demonstrates that monitoring for the induction of specific antibodies during clinical trials is an important issue for therapeutic proteins.