Production and immunohistochemical characterization of monoclonal antibodies directed against renal basement membranes of rats

Abstract

Basement membranes were separated from rat glomeruli and purified by mild procedures, which led to a highly enriched basement membrane fraction. Here, the production and characterization of five monoclonal antibodies against tubular and glomerular basement membranes are described. These antibodies were analyzed immunohistochemically on frozen sections of rat, bovine, and human kidneys as well as on rat embryos. One monoclonal antibody (BM 0 11) exclusively recognized the glomerular basement membranes, another one (BM 0 VII) bound to tubular basement membranes and to Bowman's capsule. Three antibodies (BM 0 IV, BM M 11, BM M 111) recognized their antigens in both glomerular and tubular basement membranes as well as in mesangial cells. The BM 0 I1 antibody showed a stringent species specificity and bound only to glomerular basement membranes of the rat. The other four antibodies cross-reacted with human and bovine glomerular basement membrane and mesangial antigens; they also bound to other tissues in the developing rat embryo. Antibody binding to specific purified components of the basement membranes such as collagen type IV, laminin, heparan sulphate proteoglycan, and fibronectin was investigated by enzyme-linked immunosorbent assay (ELISA). None of these antibodies reacted with any of these known basement membrane components, indicating that the antibodies may serve as useful tools in future investigations of so far unidentified components of basement membranes.