Abstract
Quantitation of gluten in gluten-free products is a great challenge as it is hindered by several factors including the lack of certified reference materials. To resolve this problem, our research group, in cooperation with other international experts, started a series of experiments with the goal of the production of a suitable gluten reference material. As a part of this research, several different wheat cultivars and their isolated gluten proteins were characterized by different methods, including enzyme-linked immunosorbent assay (ELISA). However, we need to know the performance of the ELISA methods used for this special area of research. During the present work we investigated the accuracy and precision of two different ELISA methods for our own laboratory conditions and special sample matrices (wheat flours and gliadin isolate). We have found that the tested performance characteristics of the methods seem to be appropriate on a case-by-case basis, but the long-term measurement uncertainty is higher, which makes it difficult to evaluate the results obtained with the ELISA method for these types of samples.
Highlights
Quantitation of gluten in gluten-free products is a great challenge as it is hindered by several factors including the lack of certified reference materials
The widely used standard-like material for gluten analysis is the PWG-gliadin. It is an isolated protein fraction from 28 European wheat cultivars with good solubility, homogeneity, and stability properties (VAN ECKERT et al, 2006). It was proposed for approval as a certified reference material (RM), but it does not meet some of the RM requirements, such as reproducibility of production (DIAZ-AMIGO & POPPING, 2013)
We must know if this is the uncertainty of the method, or can we identify factors that affect the performance of enzyme-linked immunosorbent assay (ELISA) methods? We determined and compared the extents of these effects and tried to identify the possible causes, as well
Summary
Quantitation of gluten in gluten-free products is a great challenge as it is hindered by several factors including the lack of certified reference materials. The determination of gluten is further complicated, because the protein content and composition of cereals vary depending on genetic (species, varieties) and environmental factors (harvest year and agricultural practices) As a result, these factors affect the relative amount of epitopes that are recognized by antibodies commonly used in gluten ELISA test kits (HAJAS et al, 2018; SCHOPF & SCHERF, 2018). The speciality of our RM development work is that we had to investigate different matrices with high gluten protein contents, like wheat flours, gluten or gliadin isolates As these types of samples are not included in the intended use of the ELISA methods, it is very important to understand their performance characteristics for this application. It is possible to include ELISA methods for gluten RM development purposes
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