GnRH complementary peptide antibodies: outcome in GnRH receptor immunoanalysis.
The aim of this study was to obtain gonadoptropin-releasing hormone (GnRH) receptor antibodies of high affinity for receptor immunoanalysis. According to the complementary peptide theory, complementary nucleic acid segments encode for the hormone ligand and the receptor binding site, respectively. On this premise, we used as immunogen, GnRH complementary peptide [N-terminal]Ser-Arg-Ala-Gln-Ser-Ile-Gly-Pro-Val-Leu conjugated with a carrier protein. High antibody titers were obtained in rabbits, rats and mice. Our antisera recognized the hydrophobic middle part of the GnRH complementary peptide. A band of protein with a molecular weight similar to that of the GnRH receptor (60 kDa) was specifically detected by immunoblot of solubilized rat pituitary membranes with the highest titering rabbit antiserum. In bioassays on sheep pituitary cells in vitro, some antisera inhibit basal or GnRH-induced LH secretion. In order to elicit antibodies of high affinity, we used a selective receptor assay on rat brain and pituitary sections where the ligand was the labeled agonist Des-Gly10-D-Ala6 GnRH. None of the highest titering antisera prevented the binding of such a high affinity ligand. The complementary peptide approach thus appears not to be optimal for obtaining high affinity antibodies against the GnRH receptor binding site.
Néri
C
C
Laboratoire de Cancérologie Expérimentale, Faculté de Médicine Nord, Marseille.
Ban
E
E
Taragnat
C
C
Caldani
M
M
Haour
F
F
Calas
B
B
Martin
P M
PM
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D001665:Binding Sites; D000939:Epitopes / Q000737:chemistry; D007987:Gonadotropin-Releasing Hormone / Q000276:immunology*; D006801:Humans; D007120:Immunochemistry; D066298:In Vitro Techniques; D007986:Luteinizing Hormone / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000494:pharmacology; D010902:Pituitary Gland / Q000187:drug effects / Q000378:metabolism; D011966:Receptors, LHRH / Q000187:drug effects / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D010455:Peptides; D011966:Receptors, LHRH; D007987:Gonadotropin-Releasing Hormone; D007986:Luteinizing Hormone