Evidence map›Paper›PMID 40385697›Full record

ArticleJournal of mass spectrometry and advances in the clinical lab2025

Quantification of glucagon and oxyntomodulin by protein precipitation-immunoaffinity enrichment-LC-MS/MS.

Jessica O Becker, Sara K Shijo, Huu-Hien Huynh, Katrina L Forrest, Michael J MacCoss, Michelle A Emrick, Elisha Goonatilleke, Andrew N Hoofnagle

Abstract read
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Article in Journal of mass spectrometry and advances in the clinical lab, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

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0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

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Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

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5 · Who and what money

Authors and funding

8 authors.

Jessica O BeckerDepartment of Laboratory Medicine and Pathology, USA.
Sara K ShijoDepartment of Laboratory Medicine and Pathology, USA.
Huu-Hien HuynhDepartment of Laboratory Medicine and Pathology, USA.
Katrina L ForrestDepartment of Laboratory Medicine and Pathology, USA.
Michael J MacCossDepartment of Genome Sciences, USA.
Michelle A EmrickDepartment of Laboratory Medicine and Pathology, USA.
Elisha GoonatillekeDepartment of Laboratory Medicine and Pathology, USA.
Andrew N HoofnagleDepartment of Laboratory Medicine and Pathology, USA.

Funding

PILOT STUDY--CLINICAL NUTRITION RESEARCHP30DK035816 · NIDDK · UNIVERSITY OF WASHINGTON · PI Ellen A Schur · 1986 to 2026
$30.4M
Quantifying proteins in plasma do democratize personalized medicine for patients with type 1 diabetesU01DK137097 · NIDDK · UNIVERSITY OF WASHINGTON · PI ANDREW N HOOFNAGLE, Michael MacCoss · 2023 to 2026
$3.4M
NIDDK NIH HHS P30 DK035816NIDDK NIH HHS U01 DK137097
6 · The paper itself

Abstract

Introduction: Glucagon and oxyntomodulin are peptide hormones differentially released from proglucagon that function in regulating blood glucose. Their overlapping amino acid sequences make the development of specific immunoassays difficult, but the specificity of liquid chromatography-tandem mass spectrometry can be used to distinguish the peptides. We aimed to develop a sensitive and specific mass spectrometric assay that uses non-proprietary reagents and normal-flow liquid chromatography in the simultaneous quantification of both analytes. Methods: Bulk plasma proteins were precipitated in ethanol/ammonium hydroxide. Analytes were enriched using monoclonal antibodies generated in-house and analyzed using liquid chromatography-tandem mass spectrometry. A glucagon calibration material was sourced commercially and characterized for purity and concentration by high-performance liquid chromatography-ultraviolet detection and amino acid analysis. Single-point calibration was used to minimize between-day variability. Results: The novel antibodies performed acceptably (peptide recovery 45-59 %). The assay was precise (<13 %CV) and linear over the range of 1.3-14.7 pM and 1.1-13.7 pM for glucagon and oxyntomodulin, respectively. The glucagon calibration material concentration was determined to be 1.596 mg/g. Tube-type studies supported the use of protease inhibitor tubes at the time of blood draw. Patients with type 1 diabetes had lower concentrations of glucagon when maintained on an insulin pump, but not with injectable insulin. Conclusion: We have validated a method with a highly detailed standard operating procedure. We have characterized calibration materials to help maintain accuracy and achieve between-day and between-laboratory harmonization. The assay will be beneficial in better understanding α-cell health and glycemic control in diabetes and other diseases.

Indexed as

Antibody enrichmentDiabetesGlucagonLiquid chromatography-tandem mass spectrometryOxyntomodulin

Identifiers

PMID40385697
PMCPMC12083559

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LicenceCC BY-NC-ND
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.