[Isolation and characteristics of monoclonal antibodies to the external domain of the EGF receptor in human A431 epidermoid carcinoma].
The monoclonal antibody to the epidermal growth factor (EGF) receptor was generated after fusion of PAI myeloma cells with immunized BALB/c mouse spleen cells, using intact A431 epidermoid carcinoma cells as an immunogen. The antibody, denoted 5A9, is an IgG, which recognizes a protein with molecular mass 170 kDa during immunoblot analysis, immunoprecipitates phosphoprotein with molecular mass 170 kDa from the membrane preparations of A431 cells, and, according to immunofluorescence experiments, is distributed in the cell similar to the EGF-rhodamine conjugate. It is concluded that the produced antibodies are specific to EGF-receptor. At the same time the 5A9 (50 nM) do not compete with EGF for binding with high and low affinity receptors. They fail to induce internalization of the EGF-receptor and do not exert influence on intracellular degradation of EGF-receptor. Monoclonal antibodies 5A9 are also unable to inhibit the EGF-induced protein kinase activity of the receptor and do not stimulate protein kinase activity by themselves. Thus, the prepared monoclonal antibodies can be used to register the EGF-receptor cellular localization without affecting biologic activity of the receptor.
Sorokin
A B
AB
Nesterov
A M
AM
Sorkin
A D
AD
Ignatova
T N
TN
Galaktionov
K I
KI
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000302:isolation & purification*; D000918:Antibody Specificity; D002294:Carcinoma, Squamous Cell / Q000032:analysis / Q000276:immunology*; D002460:Cell Line; D004705:Endocytosis; D000939:Epitopes / Q000032:analysis; D066246:ErbB Receptors / Q000032:analysis / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007114:Immunization / Q000379:methods; D015151:Immunoblotting; D007457:Iodine Radioisotopes; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007457:Iodine Radioisotopes; D066246:ErbB Receptors
Poluchenie i kharakteristika monoklonal'nykh antitel k naruzhnomu domenu retseptora EFR épidermoidnoĭ kartsinomy cheloveka A431.