New monoclonal antibodies directed against the propart segment of human prorenin as a tool for the exploration of prorenin conformation.
Six monoclonal antibodies (MAbs) directed against human prorenin were produced by immunizing BALB/c mice with a peptide corresponding to the sequence (-17 to +9) of prorenin. The new MAbs were screened for their ability to first bind to the immobilized peptide and then to prorenin previously captured by an anti-total renin MAb. The specificity of the MAbs was confirmed by the total lack of binding to active renin. Using BIAcore technology, equilibrium affinity constants of the MAbs were determined and ranged from 3.2 x 10(8) to 5.7 x 10(9) l/mol. Immunoradiometric assays (IRMA) for prorenin were performed using the anti-total renin MAb and the anti-prorenin MAbs. The best results were obtained when an anti-prorenin MAb was immobilized and the anti-total renin MAb was used as tracer in a one-step procedure. Moreover, the signal was significantly increased by the presence of the renin inhibitor SR 43845 suggesting that the inhibitor-induced conformational change of prorenin could be detected by the MAbs.
Cohen
P
P
Sanofi Recherche, Montpellier, France.
Simon
D
D
Badouaille
G
G
Mani
J C
JC
Portefaix
J M
JM
Pau
B
B
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D002459:Cell Fusion; D004151:Dipeptides / Q000494:pharmacology; D004792:Enzyme Precursors / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D011487:Protein Conformation; D011863:Radioimmunoassay; D011994:Recombinant Proteins; D012083:Renin / Q000737:chemistry / Q000276:immunology*; D000092502:Renin Inhibitors
D000911:Antibodies, Monoclonal; D004151:Dipeptides; D004792:Enzyme Precursors; D010455:Peptides; D011994:Recombinant Proteins; D012083:Renin; D000092502:Renin Inhibitors; C060501:SR 43845