Interaction of a monoclonal antibody against hEGF with a receptor site for EGF.
Epidermal growth factor (EGF) has been detected by radioimmunoassay (RIA) in different body fluids such as serum, amniotic fluid, and urine. Human tumor tissues with EGF receptors (EGF-Rc) may be saturated with EGF, which may be of prognostic value. An RIA was envisaged to measure human epidermal growth factor (hEGF) levels using EGF-Rc as capture agent and a monoclonal antibody anti-hEGF (MAb anti-hEGF) labeled with 125Iodine as a marker for this binding. The purpose of this work was to study the feasibility of MAb anti-hEGF to detect the receptor binding sites and to investigate the interaction between MAb anti-hEGF and the EGF-Rc. Various binding experiments were performed to study possible interference and interactions in the complex MAb anti-hEGF and the receptor. Affinity constants were determined by means of Scatchard plot analysis to interpret the complex stability challenged with other compounds for a better understanding of the interaction process. Binding constants were of the same order for all the ligands tested separately involving the EGF-Rc, but were significantly higher (t = 15.7, p < 0.05) for hEGF in its binding to MAb anti-hEGF. It was possible with equilibrium studies and competition experiments to evaluate the interaction of EGF and MAb anti-hEGF with the EGF receptor. This observation makes the MAb anti-hEGF a potential tracer for the quantitation of receptors in vitro, and possibly for the detection of membrane receptors on tumor cells in vivo.
Valente
S
S
Radiopharmacy Department, Nuclear Research Center, Faculty of Sciences, Montevideo, Uruguay.
Souto
B
B
Balter
H
H
Welling
M M
MM
Román
E
E
Robles
A
A
Pauwels
E K
EK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002462:Cell Membrane / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000276:immunology*; D066246:ErbB Receptors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D051379:Mice; D011786:Quality Control; D011994:Recombinant Proteins; D013363:Submandibular Gland / Q000378:metabolism
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D066246:ErbB Receptors