title: Investigation of the solid- and solution-phase binding reactivities of continuous epitopes recognized by polyclonal guinea-pig anti-recombinant bovine growth hormone antisera.
abstract: We have used the technique of multiple pin peptide synthesis to identify three major continuous epitopes in the recombinant bovine (rb) GH molecule. We have synthesized these peptides, residues 24-40, 139-152 and 179-189, as N-terminally acetylated, C-terminal amides and confirmed their reactivity in a standard solid-phase ELISA. Subsequently, for epitope 139-152, we have synthesized a peptide affinity column and used this to isolate antibodies with this epitope specificity from whole antiserum. In addition, we demonstrate that under native conditions in a liquid phase RIA, these antibodies will precipitate [125I]rbGH. Further, peptide 139-152 itself also cross-reacts in an rbGH RIA inhibiting binding by up to 20%. Our data suggest that during the immune response to rbGH in guinea-pigs a substantial part of the B-cell response is directed to the 139-152 region and that this part of the protein is a native epitope.
authors.affiliation: Hannah Research Institute, Ayr, UK.
authors.forename: J
authors.initials: J
authors.lastname: Beattie
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D000939:Epitopes / Q000276:immunology*; D005260:Female; D013006:Growth Hormone / Q000276:immunology*; D006168:Guinea Pigs; D007106:Immune Sera / Q000276:immunology*; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D012996:Solutions
chemical_list: D000939:Epitopes; D007106:Immune Sera; D010446:Peptide Fragments; D011994:Recombinant Proteins; D012996:Solutions; D013006:Growth Hormone