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

Nicolai Jacob Wewer Albrechtsen; Ali Asmar; Frederik Jensen; Signe Törang; Lene Simonsen; Rune Ehrenreich Kuhre; Meena Asmar; Simon Veedfald; Astrid Plamboeck; Filip K Knop; Tina Vilsboll; Sten Madsbad; Michael A Nauck; Carolyn F Deacon; Jens Bulow; Jens J Holst; Bolette Hartmann

doi:10.1152/ajpendo.00005.2017

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

Nicolai Jacob Wewer Albrechtsen, Ali Asmar, Frederik Jensen, Signe Törang, Lene Simonsen, Rune Ehrenreich Kuhre, Meena Asmar, Simon Veedfald, Astrid Plamboeck, Filip K Knop, Tina Vilsboll, Sten Madsbad, Michael A Nauck, Carolyn F Deacon, Jens Bulow, Jens J Holst, Bolette Hartmann

12 Citations (Scopus)

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 non-amidated 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.

Original language	English
Journal	A J P: Endocrinology and Metabolism (Online)
Pages (from-to)	e284-e291
Number of pages	7
ISSN	1522-1555
DOIs	https://doi.org/10.1152/ajpendo.00005.2017
Publication status	Published - Sept 2017

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Journal Article

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Wewer Albrechtsen, N. J., Asmar, A., Jensen, F., Törang, S., Simonsen, L., Kuhre, R. E., Asmar, M., Veedfald, S., Plamboeck, A., Knop, F. K., Vilsboll, T., Madsbad, S., Nauck, M. A., Deacon, C. F., Bulow, J., Holst, J. J., & Hartmann, B. (2017). A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite. A J P: Endocrinology and Metabolism (Online), e284-e291. https://doi.org/10.1152/ajpendo.00005.2017

Wewer Albrechtsen, NJ, Asmar, A, Jensen, F, Törang, S, Simonsen, L, Kuhre, RE, Asmar, M, Veedfald, S, Plamboeck, A, Knop, FK , Vilsboll, T , Madsbad, S, Nauck, MA, Deacon, CF, Bulow, J, Holst, JJ & Hartmann, B 2017, 'A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite', A J P: Endocrinology and Metabolism (Online), pp. e284-e291. https://doi.org/10.1152/ajpendo.00005.2017

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title = "A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite",

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 non-amidated 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 = "Journal Article",

author = "{Wewer Albrechtsen}, {Nicolai Jacob} and Ali Asmar and Frederik Jensen and Signe T{\"o}rang and Lene Simonsen and Kuhre, {Rune Ehrenreich} and Meena Asmar and Simon Veedfald and Astrid Plamboeck and Knop, {Filip K} and Tina Vilsboll and Sten Madsbad and Nauck, {Michael A} and Deacon, {Carolyn F} and Jens Bulow and Holst, {Jens J} and Bolette Hartmann",

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doi = "10.1152/ajpendo.00005.2017",

language = "English",

pages = "e284--e291",

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T1 - A sandwich ELISA for measurement of the primary glucagon-like peptide-1 metabolite

AU - Wewer Albrechtsen, Nicolai Jacob

AU - Asmar, Ali

AU - Jensen, Frederik

AU - Törang, Signe

AU - Simonsen, Lene

AU - Kuhre, Rune Ehrenreich

AU - Asmar, Meena

AU - Veedfald, Simon

AU - Plamboeck, Astrid

AU - Knop, Filip K

AU - Vilsboll, Tina

AU - Madsbad, Sten

AU - Nauck, Michael A

AU - Deacon, Carolyn F

AU - Bulow, Jens

AU - Holst, Jens J

AU - Hartmann, Bolette

PY - 2017/9

Y1 - 2017/9

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 non-amidated 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 non-amidated 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

SN - 1522-1555

SP - e284-e291

JO - A J P: Endocrinology and Metabolism (Online)

JF - A J P: Endocrinology and Metabolism (Online)

ER -

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

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