A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite
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A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite. / Wewer Albrechtsen, Nicolai J; Asmar, Ali; Jensen, Frederik; Törang, Signe; Simonsen, Lene; Kuhre, Rune E; Asmar, Meena; Veedfald, Simon; Plamboeck, Astrid; Knop, Filip K; Vilsbøll, Tina; Madsbad, Sten; Nauck, Michael A; Deacon, Carolyn F; Bülow, Jens; Holst, Jens J; Hartmann, Bolette.
In: American Journal of Physiology: Endocrinology and Metabolism, Vol. 313, No. 3, 01.09.2017, p. E284-E291.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite
AU - Wewer Albrechtsen, Nicolai J
AU - Asmar, Ali
AU - Jensen, Frederik
AU - Törang, Signe
AU - Simonsen, Lene
AU - Kuhre, Rune E
AU - Asmar, Meena
AU - Veedfald, Simon
AU - Plamboeck, Astrid
AU - Knop, Filip K
AU - Vilsbøll, Tina
AU - Madsbad, Sten
AU - Nauck, Michael A
AU - Deacon, Carolyn F
AU - Bülow, Jens
AU - Holst, Jens J
AU - Hartmann, Bolette
N1 - Copyright © 2017 the American Physiological Society.
PY - 2017/9/1
Y1 - 2017/9/1
N2 - 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.
AB - 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.
KW - Journal Article
U2 - 10.1152/ajpendo.00005.2017
DO - 10.1152/ajpendo.00005.2017
M3 - Journal article
C2 - 28420649
VL - 313
SP - E284-E291
JO - American Journal of Physiology - Endocrinology and Metabolism
JF - American Journal of Physiology - Endocrinology and Metabolism
SN - 0193-1849
IS - 3
ER -
ID: 182973352