Preparation and characterization of monoclonal antibodies specific for human transforming growth factor alpha
Sorvillo, J.M.; McCormack, E.S.; Yanez, L.; Valenzuela, D.; Reynolds, F.H.
Oncogene 5(3): 377-386
1990
ISSN/ISBN: 0950-9232 PMID: 1690379 Document Number: 360238
A series of monoclonal antibodies (mAbs) against transforming growth factor .alpha. (TGF.alpha.) have been produced. The generation of these reagents, as well as their biochemical and immunochemical characterization is described. TGF.alpha. peptides, mutant recombinant TGF.alpha. proteins and two-site immunoradiometric assays were used to identify the epitopes recognized by each antibody. This approach has allowed the specific localization of immunodominant domains on the molecule. Certain mAbs were found to be useful for selected procedures. mAb 134A-2B3 was used for immunoblotting both the precursor and mature forms of TGF.alpha. from conditioned media of tumor cells. One mAb 189-2130.1, which reacted with the carboxyl terminal seventeen amino acids, was able to block TGF.alpha. binding to the EGF receptor. mAb 213-4.4 was used for immunohistochemical detection of TGF.alpha. in fixed tumor cells. mAbs 137-178 and 134A-2B3 were used to develop a two-site immunoradiometric immunoassay which was sensitive to 1 ng ml-1 and detected TGF.alpha. from a variety of tumor cells. A series of mAbs such as these could prove useful in studying the biochemical properties as well as the immunochemical localization of TGF.alpha. in normal tissues and tumors.