A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite
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Nicolai J Wewer Albrechtsen, Ali Asmar, Frederik Jensen, Signe Törang, Lene Simonsen, Rune E Kuhre, Meena Asmar, Simon Veedfald, Astrid Plamboeck, Filip K Knop, Tina Vilsbøll, Sten Madsbad, Michael A Nauck, Carolyn F Deacon, Jens Bülow, Jens J Holst, Bolette Hartmann
Glucagon-like peptide-1 (GLP-1) is an incretin hormone secreted from the gastrointestinal tract. It is best known for its glucose-dependent insulinotropic effects. GLP-1 is secreted in its intact (active) form (7-36NH2) but is rapidly degraded by the dipeptidyl peptidase 4 (DPP-4) enzyme, converting >90% to the primary metabolite (9-36NH2) before reaching the targets via the circulation. Although originally thought to be inactive or antagonistic, GLP-1 9-36NH2 may have independent actions, and it is therefore relevant to be able to measure it. Because reliable assays were not available, we developed a sandwich ELISA recognizing both GLP-1 9-36NH2 and nonamidated GLP-1 9-37. The ELISA was validated using analytical assay validation guidelines and by comparing it to a subtraction-based method, hitherto employed for estimation of GLP-1 9-36NH2 Its accuracy was evaluated from measurements of plasma obtained during intravenous infusions (1.5 pmol × kg(-1) × min(-1)) of GLP-1 7-36NH2 in healthy subjects and patients with type 2 diabetes. Plasma levels of the endogenous GLP-1 metabolite increased during a meal challenge in patients with type 2 diabetes, and treatment with a DPP-4 inhibitor fully blocked its formation. Accurate measurements of the GLP-1 metabolite may contribute to understanding its physiology and role of GLP-1 in diabetes.
|Tidsskrift||American Journal of Physiology: Endocrinology and Metabolism|
|Status||Udgivet - 1 sep. 2017|