Vitiligo is considered an autoimmune disorder due to the generation and presence of autoantibodies directed against melanocyte antigens in the patients' sera. In the present study we point towards a newly defined autoantigen in vitiligo, the enzyme tyrosinase, which participates in the process of melanogenesis. Anti‐tyrosinase antibodies were detected in the sera of seven patients with diffuse and 11 patients with localized vitiligo. Employing solid‐phase ELISA to mushroom tyrosinase, we found that patients with diffuse vitiligo had significantly higher titres of IgG anti‐tyrosinase autoantibodies than patients with localized disease or healthy subjects. These anti‐tyrosinase autoantibodies have relatively high functional affinity to tyrosinase and can be recovered from vitiligo patients’ sera by affinity purification. The anti‐tyrosinase antibodies do not cross‐react with other enzymes recognized as autoantigens in different autoimmune disorders and the autoantibodies do not block the enzymatic activity of tyrosinase, indicating that they are not reacting with the catalytic site of the enzyme. These data point to tyrosinase as an autoantigen in vitiligo and suggest that anti‐tyrosinase titres can serve as a marker for disease activity.