Preparation and properties of monoclonal and polyclonal antibodies to mouse epidermal growth factor (EGF) receptors: evidence for cryptic EGF receptors in embryonal carcinoma cells.
Monoclonal antibodies to mouse epidermal growth factor (EGF) receptor were prepared by the immunization of rats with receptor glycoprotein purified from mouse liver by affinity chromatography on immobilized EGF. Purified mouse EGF receptor retained EGF-inducible autophosphorylating activity and was antigenic in rats and rabbits. The monoclonal antibodies cross react very poorly with human EGF receptor, while polyclonal rabbit antibodies immunoprecipitate human, rat and mouse EGF receptor equally well. The rabbit antibody blocks EGF binding to mouse fibroblast cells and, at 20-fold higher concentrations, stimulates uptake of tritiated thymidine into DNA. This indicates that antibodies bind at or close to the EGF-binding site and can mimic the effects of the growth factor. None of the monoclonals bind at the EGF site of the receptor. Immunoprecipitation, immunoblotting, 125I-EGF cross linking, 125I-surface labelling, immunohistochemistry and autophosphorylation techniques were used to delineate the basis for the induction of EGF receptors when OC15 embryonal carcinoma (EC) cells differentiate into endodermal derivatives (END). EGF-stimulated autophosphorylation of a 170 X 10(3) Mr protein in solubilized OC15 EC cells is readily detectable, although intact EC cells do not bind or respond to EGF by all other tests. The results suggest that cryptic EGF receptors are present in EC stem cells, a finding with implications in development.
Weller
A
A
Cancer Research Center, La Jolla Cancer Research Foundation, CA 92037.
Meek
J
J
Adamson
E D
ED
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001345:Autoradiography; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism; D054278:Embryonal Carcinoma Stem Cells; D066246:ErbB Receptors / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D051379:Mice; D014411:Neoplastic Stem Cells / Q000276:immunology*; D013936:Thymidine / Q000378:metabolism
D000906:Antibodies; D000911:Antibodies, Monoclonal; D004247:DNA; D066246:ErbB Receptors; D013936:Thymidine