Four unique monoclonal antibodies to the putative receptor binding domain of erythropoietin inhibit the biological function of the hormone.
We have produced a series of monoclonal antibodies (MoAbs) to amino acid region 99-129 of human erythropoietin (Epo) that distinguish unique structural features within this putative receptor binding domain of the hormone. The MoAbs recognize denatured Epo with widely different sensitivities on a Western blot and differentially bind to native Epo in solution. In addition, three of the four MoAbs neutralize the biological activity of Epo in a concentration-dependent fashion in vitro. Neutralization was measured both by inhibition of Epo-induced differentiation in Rauscher murine erythroleukemia cells and by inhibition of Epo-induced proliferation in normal murine splenic erythroid precursors. Characterization of the structural epitopes recognized by each of these four reagent MoAbs should provide us with important information concerning the requirements for hormone-receptor interaction.
Feldman
L
L
Department of Medicine, New England Deaconess Hospital, Harvard Medical School, Boston, Massachusetts 02215.
Heinzerling
R
R
Hillam
R P
RP
Chern
Y E
YE
Frazier
J G
JG
Davis
K L
KL
Sytkowski
A J
AJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000502:physiology*; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D004921:Erythropoietin / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D007457:Iodine Radioisotopes; D004915:Leukemia, Erythroblastic, Acute / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests; D011956:Receptors, Cell Surface / Q000276:immunology* / Q000378:metabolism / Q000502:physiology*; D017467:Receptors, Erythropoietin; D013154:Spleen / Q000166:cytology / Q000648:ultrastructure; D014407:Tumor Cells, Cultured / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007457:Iodine Radioisotopes; D011956:Receptors, Cell Surface; D017467:Receptors, Erythropoietin; D004921:Erythropoietin