Production of anti-glucagon sera with a C-terminal fragment of pancreatic glucagon.
The C-terminal region-sepcific anti-glucagon sera were raised in rabbits using as immunogen, and conjugate of BSA and a C-terminal fragment of pancreatic glucagon. The hapten was prepared by trypsin digestion of the glucagon, which was proved to be a 1:3 mixture of glucagon (18--29) and (19--29). Six rabbits were immunized by subcutaneous injection of an emulsion of the conjugate with complete Freund's adjuvant and five of the rabbits produced antibodies to the glucagon (GC-1, GC-2, GC-3, GC-5 and GC-6). For comparison, rabbit antisera were also produced against glucagon polymer (GA-10) and syrupy glucagon fibrils (PGA-2). All these antisera as well as the pancreatic glucagon-specific antiserum 30 K were characterized with dog gut-extract (gut-GLI) and glucagon-related peptide fragments in the radioimmunoassay systems. The assay systems utilized 125 I-monosubstituted pancreatic glucagon as tracer and human mono-component glucagon as standard. All sera of the GC-series crossreacted with the dog gut-extract very weakly and antisera GC-5 and GC-6 exhibited the lowest crossreactivities with the extract, which were shown to be as low as that of 30k. Characterization of the antiserum GC-5 with purified glucagon related fragments indicated that the major antigenic determinant located exactly in the C-terminal region of glucagon. The present results clearly showed high efficiency of the use of the glucagon C-terminal fragment as hepatenic immunogen in obtaining the C-terminal region-specific, i.e., pancreatic glucagon-specific antisera.
D000818:Animals; D000917:Antibody Formation; D003429:Cross Reactions; D004285:Dogs; D000939:Epitopes; D005934:Glucagon / Q000032:analysis / Q000276:immunology*; D006801:Humans; D007106:Immune Sera; D010454:Peptide Termination Factors / Q000276:immunology*; D011817:Rabbits; D011863:Radioimmunoassay; D013552:Swine; D014020:Tissue Extracts / Q000276:immunology
D000939:Epitopes; D007106:Immune Sera; D010454:Peptide Termination Factors; D014020:Tissue Extracts; D005934:Glucagon