Specific Glucagon Assay System Using a Receptor-Derived Glucagon-Binding Peptide Probe

Glucagon is a peptide hormone secreted by pancreatic alpha cells which elevates blood glucose and plays a critical role in diabetes onset and homeostasis. The accurate assessment of glucagon concentration is challenging due to its structural similarity with other hormones, causing cross-reactivity in antibody-based methods. Rapid and specific glucagon detection is essential, particularly during hypoglycemia. This study aimed to develop glucagon-specific probes combining high specificity, rapid detection, and ease of operation. We designed novel peptide-based probes by screening glucagon-binding peptides from the glucagon receptor sequence using a peptide array method. This strategy, based on receptor amino acid sequences, can be applied to the identification of binding peptides for other hormones, expanding its potential utility. The screened peptides were conjugated with fluorescent dyes to create probes enabling detection within 30 min. The developed probes demonstrated superior specificity for glucagon relative to similar sequence analogs compared with conventional antibody-based methods, with detection limits in the nanomolar range. This study represents a proof-of-concept approach for rapid and highly specific glucagon detection. However, further optimization of probe sensitivity and validation under physiological conditions will be required before clinical or diagnostic application. These improvements in the probe's properties will enable the reliable blood glucagon detection and accurate diagnostic assessment of diabetes-related diseases.