Characteristics of antibodies to the epidermal growth factor receptor-kinase.
Polyclonal antibodies to different antigenic forms of the epidermal growth factor (EGF) receptor-kinase from human A-431 cells have been produced, and their properties have been characterized and compared. Biochemically active receptor-kinase purified by affinity chromatography was employed as one type of antigen. Denatured receptor-kinase prepared by sodium dodecyl sulfate-gel electrophoresis of the affinity-purified receptor was used as the second type of antigen. Animals immunized with either type of antigen produced antibody capable of immunoprecipitating the receptor-kinase molecule. Antibodies produced in response to the biochemically active antigenic form of the receptor-kinase are capable of blocking 125I-EGF binding to the receptor and inhibited EGF-stimulated biological responses. These antisera are not species specific in their ability to inhibit growth-factor binding to the EGF receptor of various mammalian cells. However, these rabbit antisera were unable to inhibit 125I-EGF binding to rabbit cells. Although antisera produced in response to the denatured receptor-kinase molecule are not able to block 125I-EGF binding or EGF-stimulated biological responses, they are particularly efficient for the immunoprecipitation of solubilized 125I-EGF:receptor complexes. None of the antisera contain antibodies capable of interfering with basal receptor-kinase phosphorylation activity. Although each of the antisera immunoprecipitated this kinase activity, none of the antisera contained antibody which served as a phosphorylation substrate for the EGF receptor-kinase in contrast to the immunoglobulins present antisera to the src gene product of the Rous sarcoma virus.
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000937:Antigen-Antibody Reactions; D011232:Chemical Precipitation; D004247:DNA / Q000096:biosynthesis; D066246:ErbB Receptors; D007120:Immunochemistry; D051379:Mice; D010766:Phosphorylation; D011494:Protein Kinases / Q000276:immunology*; D011817:Rabbits
D000906:Antibodies; D004247:DNA; D011494:Protein Kinases; D066246:ErbB Receptors