Production of glucagon antibodies and their role in metabolism and immunoassay of glucagon

SummaryThe antibody content of sera from rabbits treated intermittently with free glucagon or glucagon conjugated with ovalbumin was measured at intervals during a 2-year period. Circulating nonprecipitating antibody, capable of binding glucagon-I131, was detectable within one month, and was still detectable after 4 to 5 month periods during which no antigen was administered. Glucagon-I131 injected into an immunized rabbit was rapidly bound to antibody, remained chromatographically intact and was only slowly cleared from the circulation. Glucagon-I131 could be dissociated from its antibody at acid pH. The preferential salt precipitation technic used for immunochemical assay of insulin proved applicable to measurement of minute amounts of glucagon.The authors are indebted to Dr. Otto K. Behrens, Eli Lilly & Co. for supplies of recrystallized glucagon and for tests of antibody inhibition of glucagon activity in cats. We wish to acknowledge the technical assistance of Miss Angela Lee.