title: Monoclonal anti-gonadotropin releasing hormone (GnRH) antibodies reacting to sequence of GnRH preferentially recognize to native hormone.
abstract: Monoclonal anti-GnRH antibodies (MoAbs) P862, P778, P813 and P764 reacted optimally to native GnRH and poorly to GnRH(OH) of sequence 4-6, 7-10, 4-10 and 1-10. The heptapeptide 4-10 showed maximum reactivity amongst the four peptides tested for immunoreactivity. Sepharose 4B-GnRH(OH)4-10 (H-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-OH) column was therefore used to purify the fraction of MoAb reacting to this sequence. The affinity purified MoAbs (A-MoAbs) were further characterized for their binding to the different sequences and affinity with native GnRH. The binding, cross-reactivity and affinity characteristics of A-MoAbs were comparable with those of MoAbs. Immunoreactivity of A-MoAbs was also observed to be partly regained when GnRH(OH)1-10 was coupled to Lys, Lys-MDP or H-Ala-Ala-Thr-Lys-Pro-Arg-OH. These observations clearly demonstrate that MoAbs were neither contaminated nor were sequence specific but were directed against the conformation of the molecule.
authors.forename: V
authors.initials: V
authors.lastname: Singh
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D001667:Binding, Competitive; D002846:Chromatography, Affinity; D008968:Molecular Conformation; D010906:Pituitary Hormone-Releasing Hormones / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D010906:Pituitary Hormone-Releasing Hormones