A monoclonal antibody against human kidney gamma-glutamyl transpeptidase: Preparation, immunochemical, and immunohistochemical characterization

To perform immunohistochemical study of the distribution of γ-glutamyl transpeptidase in human organs, a highly specific antibody against the human enzyme is required. We prepared monoclonal antibody against γ-glutamyl transpeptidase from human kidney, using the hybridoma technique. The antibody was of the IgG₁ type and the light chain belonged to the κ class. The antibody reacted specifically with the 63 KD heavy subunit of the enzyme. Examination of the specificity of the antibody performed by immunohistochemical staining of human kidney sections revealed that the antigen was localized on the brush border and along the basolateral membrane of the epithelial cells of both the convoluted and the straight portions of the proximal tubule. This antibody was also reactive in several human organs other than kidney, including epididymis, prostate, seminal vesicle, pancreas, and normal liver, and in human hepatoma. These findings indicate the existence of an antigenic determinant common to human kidney and other organs. The monoclonal antibody did not crossreact with mouse, rat, guinea pig, rabbit, or pig kidney.