Internal image properties of a monoclonal auto-anti-idiotypic antibody and its binding to aldosterone receptors.
A monoclonal anti-idiotypic antibody (H10E4C9F) that interacts with the aldosterone receptors was generated using an auto-anti-idiotypic approach by immunizing a mouse with a 3-O-carboxymethyloxime of aldosterone coupled to bovine serum albumin. This antibody, an IgG1, displayed internal image properties of aldosterone and was considered as an Ab2 beta according to the following criteria. (i) H10E bound to Fab fragments of affinity-purified rabbit anti-aldosterone antibody that had high affinity for aldosterone (Kd = 5 x 10(-10) M). Binding was inhibited by aldosterone but not by estradiol. (ii) H10E inhibited [3H]aldosterone binding to rabbit polyclonal antibodies and also to murine monoclonal antibodies raised during the same fusion. Inhibition was concentration-dependent. These results are consistent with the antibody recognizing an interspecies cross-reacting epitope involved in the aldosterone combining site. (iii) The antibody could be affinity-purified on an immobilized monoclonal anti-aldosterone antibody. (iv) It inhibited [3H]aldosterone binding to rabbit kidney cytosolic aldosterone receptors but had no effect on glucocorticoid receptors. Additional evidence for the interaction of H10E with aldosterone receptors was provided by glycerol gradients analyses: the anti-idiotypic antibody displaced [3H]aldosterone and [3H]corticosterone from the native untransformed 9 S aldosterone receptor in the presence of RU 26988, a specific marker of glucocorticoid receptors. All of the above are consistent with the first successful production of a monoclonal antibody that mimics aldosterone and interacts specifically with the steroid binding domain of aldosterone receptors.
Lombes
M
M
Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Edelman
I S
IS
Erlanger
B F
BF
D000315:Adrenalectomy; D000450:Aldosterone / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D003600:Cytosol / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D007140:Immunoglobulin Fab Fragments; D007130:Immunoglobulin Idiotypes / Q000276:immunology*; D007668:Kidney / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000276:immunology* / Q000378:metabolism; D018161:Receptors, Mineralocorticoid
D000906:Antibodies; D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D007140:Immunoglobulin Fab Fragments; D007130:Immunoglobulin Idiotypes; D011965:Receptors, Glucocorticoid; D018161:Receptors, Mineralocorticoid; D000450:Aldosterone