Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.

<h4>Background</h4>Subjects suffering from coeliac disease, gluten allergy/intolerance must adopt a lifelong avoidance of gluten. Beer contains trace levels of hordeins (gluten) which are too high to be safely consumed by most coeliacs. Accurate measurement of trace hordeins by ELISA is...

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Autores principales:	Gregory J Tanner, Michelle L Colgrave, Malcolm J Blundell, Hareshwar P Goswami, Crispin A Howitt
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/5e09fff8c9a94a648888d14ab4b2ff50
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spelling	oai:doaj.org-article:5e09fff8c9a94a648888d14ab4b2ff502021-11-18T07:55:16ZMeasuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.1932-620310.1371/journal.pone.0056452https://doaj.org/article/5e09fff8c9a94a648888d14ab4b2ff502013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23509606/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Subjects suffering from coeliac disease, gluten allergy/intolerance must adopt a lifelong avoidance of gluten. Beer contains trace levels of hordeins (gluten) which are too high to be safely consumed by most coeliacs. Accurate measurement of trace hordeins by ELISA is problematic.<h4>Methods</h4>We have compared hordein levels in sixty beers, by sandwich ELISA, with the level determined using multiple reaction monitoring mass spectrometry (MRM-MS).<h4>Results</h4>Hordein levels measured by ELISA varied by four orders of magnitude, from zero (for known gluten-free beers) to 47,000 µg/mL (ppm; for a wheat-based beer). Half the commercial gluten-free beers were free of hordein by MS and ELISA. Two gluten-free and two low-gluten beers had zero ELISA readings, but contained significant hordein levels (p<0.05), or near average (60-140%) hordein levels, by MS, respectively. Six beers gave false negatives, with zero ELISA readings but near average hordein content by MS. Approximately 20% of commercial beers had ELISA readings less than 1 ppm, but a near average hordein content by MS. Several barley beers also contained undeclared wheat proteins.<h4>Conclusions</h4>ELISA results did not correlate with the relative content of hordein peptides determined by MS, with all barley based beers containing hordein. We suggest that mass spectrometry is more reliable than ELISA, as ELISA enumerates only the concentration of particular amino-acid epitopes; this may vary between different hordeins and may not be related to the absolute hordein concentration. MS quantification is undertaken using peptides that are specific and unique, enabling the quantification of individual hordein isoforms. This outlines the problem of relying solely on ELISA determination of gluten in beverages such as beer and highlights the need for the development of new sensitive and selective quantitative assay such as MS.Gregory J TannerMichelle L ColgraveMalcolm J BlundellHareshwar P GoswamiCrispin A HowittPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 2, p e56452 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Gregory J Tanner Michelle L Colgrave Malcolm J Blundell Hareshwar P Goswami Crispin A Howitt Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.
description	<h4>Background</h4>Subjects suffering from coeliac disease, gluten allergy/intolerance must adopt a lifelong avoidance of gluten. Beer contains trace levels of hordeins (gluten) which are too high to be safely consumed by most coeliacs. Accurate measurement of trace hordeins by ELISA is problematic.<h4>Methods</h4>We have compared hordein levels in sixty beers, by sandwich ELISA, with the level determined using multiple reaction monitoring mass spectrometry (MRM-MS).<h4>Results</h4>Hordein levels measured by ELISA varied by four orders of magnitude, from zero (for known gluten-free beers) to 47,000 µg/mL (ppm; for a wheat-based beer). Half the commercial gluten-free beers were free of hordein by MS and ELISA. Two gluten-free and two low-gluten beers had zero ELISA readings, but contained significant hordein levels (p<0.05), or near average (60-140%) hordein levels, by MS, respectively. Six beers gave false negatives, with zero ELISA readings but near average hordein content by MS. Approximately 20% of commercial beers had ELISA readings less than 1 ppm, but a near average hordein content by MS. Several barley beers also contained undeclared wheat proteins.<h4>Conclusions</h4>ELISA results did not correlate with the relative content of hordein peptides determined by MS, with all barley based beers containing hordein. We suggest that mass spectrometry is more reliable than ELISA, as ELISA enumerates only the concentration of particular amino-acid epitopes; this may vary between different hordeins and may not be related to the absolute hordein concentration. MS quantification is undertaken using peptides that are specific and unique, enabling the quantification of individual hordein isoforms. This outlines the problem of relying solely on ELISA determination of gluten in beverages such as beer and highlights the need for the development of new sensitive and selective quantitative assay such as MS.
format	article
author	Gregory J Tanner Michelle L Colgrave Malcolm J Blundell Hareshwar P Goswami Crispin A Howitt
author_facet	Gregory J Tanner Michelle L Colgrave Malcolm J Blundell Hareshwar P Goswami Crispin A Howitt
author_sort	Gregory J Tanner
title	Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.
title_short	Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.
title_full	Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.
title_fullStr	Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.
title_full_unstemmed	Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.
title_sort	measuring hordein (gluten) in beer--a comparison of elisa and mass spectrometry.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/5e09fff8c9a94a648888d14ab4b2ff50
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Measuring hordein (gluten) in beer--a comparison of ELISA and mass spectrometry.

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