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Showing results for tags 'elisa'.
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Einkorn Wheat Easier to Digest, Less Toxic than Durum for Celiacs
Scott Adams posted an article in Latest Research
Celiac.com 08/04/2020 - People with celiac disease need to avoid wheat. That includes ancient Einkorn wheat, even though some folks claim it's easier to digest and less toxic than Durum and other wheat strains. Recent studies suggested that gliadin proteins from the ancient diploid einkorn wheat Triticum monococcum retained a reduced number of immunogenic peptides for celiac disease patients, and is less toxic than other more recent strains of wheat. Some experiments have shown Einkorn to be more easily digested than hexaploid common wheat. But is it really, and how does it compare in terms of toxicity to people with celiac disease? For people with celiac disease, is there a difference in digestibility and toxicity between Durum and Einkorn Wheat? Researchers say there is. A team of researchers recently set out to compare the immunological properties of gliadins from two Triticum monococcum cultivars (Hammurabi and Norberto-ID331) with those of a Triticum durum cultivar (Adamello). The research team included Luigia Di Stasio, Stefania Picascia, Renata Auricchio, Serena Vitale, Laura Gazza, Gianluca Picariello, Carmen Gianfrani, and Gianfranco Mamone. They are variously affiliated with the Institute of Food Sciences, National Research Council, Avellino, Italy; and Institute of Biochemistry and Cell Biology, National Research Council, Naples, Italy; the Department of Translational Medical Science, Section of Paediatrics, European Laboratory for the Investigation of Food-Induced Diseases, University "Federico II", Naples, Italy; the CREA-Research Centre for Engineering and Agro-Food Processing, Rome, Italy. Their team compared the immunological properties of gliadins from two Einkorn strains of Triticum monococcum (Hammurabi and Norberto-ID331) with a Triticum durum strain, called Adamello. To do so, the team digested the gliadins by duplicating in vitro gastrointestinal digestion, including the brush border membrane peptidases. Competitive ELISA, based on R5 monoclonal antibody, showed that gastrointestinal digestion reduced the immunogenicity of Triticum monococcum gliadins, but had little effect on the toxic potential of Triticum durum gliadins. The team also assessed the immune stimulatory activity by detecting the IFN-γ production in gliadin-reactive T-cell lines taken from the small intestinal mucosa of HLA-DQ2+ celiac disease patients. Notably, gastrointestinal digestion sharply reduced the ability of Einkorn gliadins of both strains to activate T cells, while barely affecting the ability of Triticum durum. The team's results clearly show that digestion barely affects Triticum durum at all, while the Einkorn strains break down substantially, resulting in lower toxicity for celiac disease patients. This study does not mean that Einkorn is safe for people with celiac disease to eat, but it does present some interesting possibilities for further study. It would be interesting, for example, to see if Einkorn could be further digested with sourdough cultures, or even enzymes, to render a bread that could be safe for people with celiac disease. The possibilities are intriguing. Stay tuned for more on this and related stories. Read more at: Front Nutr. 2020 May 22;7:56.- 7 comments
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Solid Phase Plates Are Crucial to Accurate ELISA Tests
Jefferson Adams posted an article in Latest Research
Celiac.com 02/05/2020 - The enzyme-linked immunosorbent assay (ELISA) is a critical diagnostic tool for spotting or differentiating diseases, such as celiac disease, and other autoimmune conditions, and also to monitor disease progression or drug efficacy. However, accuracy is a crucial factor in such tests. Chitinase-3-like protein 1, aka CHI3L1, is a potential new culprit in inflammatory bowel disease (IBD). A team of researchers recently set out to develop an ELISA that can identify IgA and IgG autoantibodies against chitinase-3-like protein 1 (CHI3L1), in the serum of both control and disease groups. The research team included Claudia Deutschmann, Dirk Roggenbuck, Peter Schierack, and Stefan Rödigera. They are variously affiliated with the Institute of Biotechnology, Faculty Environment and Natural Sciences, Brandenburg University of Technology Cottbus-Senftenberg, Universitätsplatz 1, 01968 Senftenberg, Germany; and the Faculty of Health Sciences, Joint Faculty of the Brandenburg University of Technology Cottbus - Senftenberg, The Brandenburg Medical School Theodor Fontane, The University of Potsdam, Senftenberg, Germany. While ELISA is often used to spot the difference between diseased or healthy states, it can also be used for quantitative assessment of biomarkers, like autoantibodies. In this way, ELISA technology can help to discover and confirm new autoantibodies. Researchers have been keen to develop immunoassays that can detect diseases, such as celiac disease in the early stages. To produce an ELISA, researchers must work extra hard, and they can be forced to deal with numerous factors, including buffer systems or antigen-autoantibody interactions. This often means lots of testing to dial-in several blocking substances to yield highly accurate results. In this case, the research team developed an ELISA to detect IgA and IgG autoantibodies against chitinase-3-like protein 1 (CHI3L1), a newly identified autoantigen in inflammatory bowel disease (IBD), in the serum of 23 control subjects and 23 disease patients. The spot any issues with reproducibility, the team tested microwell plates with both PolySorp and MaxiSorp surface coatings. Among the important issues they discovered with the surface properties of the microwell plates: IgA antibody reactivity was markedly lower, since it was in the range of background noise, when measured on MaxiSorp coated plates (p < 0.0001). The IgG antibody reactivity did not change between plates, but the plate surface significantly influenced test results (p = 0.0005). The team's report, published in Heliyon highlights the properties of solid phases and their effects on the detection of autoantibodies by ELISA. They advise that using the wrong plate can cause false negative readings, which can impair the discovery of autoantibodies, such as Chitinase-3-like protein 1, aka CHI3L1. Poor accuracy can impair the ability of doctors to accurately diagnose celiac and other diseases. Read more in Heliyon. 2020 Jan; 6(1): e03270.- 1 comment
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Celiac.com Article:Solid Phase Plates Are Crucial to Accurate ELISA Tests View full article
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Celiac.com 02/17/2017 - In recent tests, researchers found that microwave treatment (MWT) of wet wheat kernels caused a striking reduction in R5-antibody-based ELISA gluten readings, reducing the readings to under 20 ppm, so that wheat could theoretically be labeled as gluten-free. However, the actual gluten content of the wheat remained unchanged. Just the test reading changed. The research team included C Gianfrani, G Mamone, B la Gatta, A Camarca, L Di Stasio, F Maurano, S Picascia, V Capozzi, G Perna, G Picariello, A Di Luccia. They are variously affiliated with the Institute of Protein Biochemistry, CNR, Naples, Italy, the Institute of Food Sciences, CNR Avellino, Italy, the Department of the Sciences of Agriculture, Food and Environment at the University of Foggia, Italy, the Institute of Food Sciences, CNR Avellino, Italy; Department of Agriculture, University of Naples, Portici (Na), Italy, the Department of Clinical and Experimental Medicine, University of Foggia, Foggia (Italy) and National Institute of Nuclear Physics, Section of Bari, Italy, and the Department of the Sciences of Agriculture, Food and Environment, University of Foggia, Italy. The failure of R5 Elisa to register gluten in MWT stands in stark contrast to analysis of gluten peptides by G12 antibody-based ELISA, mass spectrometry-based proteomics, and in vitro assay with T cells of celiac subjects, all three of which gave consistent results both before and after MWT. As to what caused the R5 Elisa to misread the MWT samples, an SDS-PAGE analysis and Raman spectroscopy showed that MWT reduced the alcohol solubility of gliadins, and altered the access of R5-antibody to the gluten epitopes. Thus, MWT neither destroys gluten nor modifies chemically the toxic epitopes, this contradicts claims that MWT of wheat kernels detoxifies gluten. This study provides evidence that R5-antibody ELISA alone is not effective to determine gluten levels in thermally treated wheat products. Gluten epitopes in processed wheat should be monitored using strategies based on combined immunoassays with T cells from celiacs, G12-antibody ELISA after proteolysis and proper molecular characterization. Source: Food Chem Toxicol. 2017 Jan 12;101:105-113. doi: 10.1016/j.fct.2017.01.010.
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Are Gluten ELISA Test Kits Wildly Inaccurate?
Jefferson Adams posted an article in Additional Concerns
Celiac.com 03/16/2016 - If you have celiac disease, particularly if you are highly sensitive to gluten exposure, you may rely on commercial ELISA test kits for gluten detection. There are a large variety of enzyme-linked immunosorbent assays (ELISAs) commercially available for gluten detection in food, including new formats and assays with antibodies against relevant gluten epitopes. But, how accurate are these test kits for gluten detection? How reliable are they for people with celiac disease? A team of researchers recently set out to evaluate the accuracy of 14 ELISA kits for gluten detection. The kits they tested cover the full range of the current commercially available ELISA test kits. The researcher team included Ilona D. Bruins Slot, Maria G. E. G. Bremer, Ine van der Fels-Klerx, with RIKILT–Wageningen UR, Wageningen, the Netherlands, and Rob J. Hamer with the Laboratory of Food Chemistry at the Wageningen University and Research Centre in Wageningen, the Netherlands. In this study, the team assessed the performance of these kits in determining gluten content in a series of relevant food matrices varying in complexity. Their results show that none of the currently available ELISA methods can accurately detect and quantify gluten in all cases. This includes the current type I method R5 as recommended by Codex Alimentarius. In the face of these results, the team is calling for urgent improvements to testing kits, and recommends focusing on competitive formats, improving extraction methods, and the detection of relevant gluten peptides. Source: Cerealchemistry.aaccnet.org; September/October 2015, Volume 92, Number 5, Pages 513-521
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