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CGRP measurements in human plasma – a methodological study

BACKGROUND: Calcitonin gene-related peptide plasma levels have frequently been determined as a biomarker for primary headaches. However, published data is often inconsistent resulting from different methods that are not precisely described in most studies. METHODS: We applied a well-proven enzyme-li...

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Autores principales: Messlinger, Karl, Vogler, Birgit, Kuhn, Annette, Sertel-Nakajima, Julika, Frank, Florian, Broessner, Gregor
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8592105/
https://www.ncbi.nlm.nih.gov/pubmed/34266288
http://dx.doi.org/10.1177/03331024211024161
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author Messlinger, Karl
Vogler, Birgit
Kuhn, Annette
Sertel-Nakajima, Julika
Frank, Florian
Broessner, Gregor
author_facet Messlinger, Karl
Vogler, Birgit
Kuhn, Annette
Sertel-Nakajima, Julika
Frank, Florian
Broessner, Gregor
author_sort Messlinger, Karl
collection PubMed
description BACKGROUND: Calcitonin gene-related peptide plasma levels have frequently been determined as a biomarker for primary headaches. However, published data is often inconsistent resulting from different methods that are not precisely described in most studies. METHODS: We applied a well-proven enzyme-linked immunosorbent assay to measure calcitonin gene-related peptide concentrations in human blood plasma, we modified parameters of plasma preparation and protein purification and used calcitonin gene-related peptide-free plasma for standard solutions, which are described in detail. RESULTS: Calcitonin gene-related peptide levels are stable in plasma with peptidase inhibitors and after deep-freezing. Calcitonin gene-related peptide standard solutions based on synthetic intercellular fluid or pooled plasma with pre-absorbed calcitonin gene-related peptide influenced the measurements but yielded both comprehensible results. In a sample of 56 healthy subjects the calcitonin gene-related peptide plasma levels varied considerably from low (<50 pg/mL) to very high (>500 pg/mL) values. After a 12-hour exposure of these subjects to normobaric hypoxia the individual calcitonin gene-related peptide levels remained stable. CONCLUSION: Buffering with peptidase inhibitors and immediate freezing or processing of plasma samples is essential to achieve reliable measurements. Individuals show considerable differences and partly high calcitonin gene-related peptide plasma levels without detectable pathological reason. Thus plasma measurements are suited particularly to follow calcitonin gene-related peptide levels in longitudinal studies. The use of data for this study was approved by the Ethics Committee of the Medical University of Innsbruck (https://www.i-med.ac.at/ethikkommission/; EK Nr: 1242/2017).
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spelling pubmed-85921052021-11-16 CGRP measurements in human plasma – a methodological study Messlinger, Karl Vogler, Birgit Kuhn, Annette Sertel-Nakajima, Julika Frank, Florian Broessner, Gregor Cephalalgia Original Articles BACKGROUND: Calcitonin gene-related peptide plasma levels have frequently been determined as a biomarker for primary headaches. However, published data is often inconsistent resulting from different methods that are not precisely described in most studies. METHODS: We applied a well-proven enzyme-linked immunosorbent assay to measure calcitonin gene-related peptide concentrations in human blood plasma, we modified parameters of plasma preparation and protein purification and used calcitonin gene-related peptide-free plasma for standard solutions, which are described in detail. RESULTS: Calcitonin gene-related peptide levels are stable in plasma with peptidase inhibitors and after deep-freezing. Calcitonin gene-related peptide standard solutions based on synthetic intercellular fluid or pooled plasma with pre-absorbed calcitonin gene-related peptide influenced the measurements but yielded both comprehensible results. In a sample of 56 healthy subjects the calcitonin gene-related peptide plasma levels varied considerably from low (<50 pg/mL) to very high (>500 pg/mL) values. After a 12-hour exposure of these subjects to normobaric hypoxia the individual calcitonin gene-related peptide levels remained stable. CONCLUSION: Buffering with peptidase inhibitors and immediate freezing or processing of plasma samples is essential to achieve reliable measurements. Individuals show considerable differences and partly high calcitonin gene-related peptide plasma levels without detectable pathological reason. Thus plasma measurements are suited particularly to follow calcitonin gene-related peptide levels in longitudinal studies. The use of data for this study was approved by the Ethics Committee of the Medical University of Innsbruck (https://www.i-med.ac.at/ethikkommission/; EK Nr: 1242/2017). SAGE Publications 2021-07-16 2021-11 /pmc/articles/PMC8592105/ /pubmed/34266288 http://dx.doi.org/10.1177/03331024211024161 Text en © International Headache Society 2021 https://creativecommons.org/licenses/by-nc/4.0/This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (https://creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Original Articles
Messlinger, Karl
Vogler, Birgit
Kuhn, Annette
Sertel-Nakajima, Julika
Frank, Florian
Broessner, Gregor
CGRP measurements in human plasma – a methodological study
title CGRP measurements in human plasma – a methodological study
title_full CGRP measurements in human plasma – a methodological study
title_fullStr CGRP measurements in human plasma – a methodological study
title_full_unstemmed CGRP measurements in human plasma – a methodological study
title_short CGRP measurements in human plasma – a methodological study
title_sort cgrp measurements in human plasma – a methodological study
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8592105/
https://www.ncbi.nlm.nih.gov/pubmed/34266288
http://dx.doi.org/10.1177/03331024211024161
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