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Aims/introduction  The aims of the present study were to investigate the performance of a novel sandwich enzyme‐linked immunosorbent assay ( ELISA ) for measuring glucagon (1–29) with monoclonal antibodies against both the C‐ and N‐terminal regions of glucagon (1–29), and to analyze the differences in plasma levels and responses of glucagon (1–29) to oral glucose loading in normal glucose tolerance ( NGT ) subjects and patients with type 2 diabetes mellitus.    Materials and Methods  The cross‐reactivity against proglucagon fragments using the  ELISA  kit and two types of conventional radioimmunoassay ( RIA ) kits was evaluated. A 75‐g oral glucose tolerance test was carried out with  NGT  subjects and patients with type 2 diabetes mellitus, and the glucagon (1–29) concentration was measured using three types of kit.    Results  The  ELISA  kit clearly had the lowest cross‐reactivity against miniglucagon (19–29) and glicentin (1–61). The oral glucose tolerance test was carried out with 30  NGT  and 17 patients with type 2 diabetes mellitus. The glucagon (1–29) levels measured by the  ELISA  kit after glucose loading were significantly higher at all time‐points in the type 2 diabetes mellitus group than in the  NGT  group. However, the glucagon (1–29) levels measured by one  RIA  kit were significantly higher in the  NGT  group, and those measured with the other  RIA  kit were approximately the same among the groups.    Conclusions  The novel sandwich  ELISA  accurately determines plasma glucagon (1–29) concentrations with much less cross‐reactivity against other proglucagon fragments than conventional  RIA  kits.

Postabsorptive hyperglucagonemia in patients with type 2 diabetes mellitus analyzed with a novel enzyme‐linked immunosorbent assay