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Am J Physiol Endocrinol Metab. 2017 Sep 1;313(3):E284-E291. doi: 10.1152/ajpendo.00005.2017. Epub 2017 Apr 18.

A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite.

Author information

1
Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
2
Novo Nordisk Foundation Center for Basic Metabolic Research, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
3
Department of Clinical Physiology and Nuclear Medicine, Bispebjerg University Hospital, Copenhagen, Denmark.
4
Center for Diabetes Research, Gentofte Hospital, University of Copenhagen, Hellerup, Denmark.
5
Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
6
Steno Diabetes Center Copenhagen, University of Copenhagen, Gentofte, Denmark.
7
Department of Endocrinology, Hvidovre University Hospital, Copenhagen, Denmark; and.
8
Division of Diabetology, Department of Medicine I, St. Josef Hospital, Ruhr University of Bochum, Bochum, Germany.
9
Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark; jjholst@sund.ku.dk.

Abstract

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.

KEYWORDS:

DPP-4; ELISA; GLP-1; GLP-1 metabolite; diabetes

PMID:
28420649
DOI:
10.1152/ajpendo.00005.2017
[Indexed for MEDLINE]
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