[Radioligand assay: method and application. II. Effect of glucagon coupling on the production of glucagon antisera with suitable binding parameters for radioimmunoassay].

Glucagon antibodies were produced in rabbits using three immunogenic glucagon conjugates. Glucagon was coupled to bovine serum albumin by difluorodinitrobenzene (DFDNB), carbodiimide (ECDI) or glutardialdehyde. Rabbits immunized with glucagon conjugated to albumin using DFDNB produced sensitive antisera for radioimmunoassay quite specific for pancreatic glucagon. The affinity constant of the best antiserum was approximately 10(11) l/mol. Antisera raised against the two other glucagon-conjugates were significantly less affine. All of these antisera showed inverse binding curves of 126J-glucagon caused by positive cooperatively in dependence upon the antigen/antibody ratio. The race of rabbits used for immunization was without influence on the immune response.