Document Detail


Quantification of gliadin levels to the picogram level by flow cytometry.
MedLine Citation:
PMID:  15648078     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: Celiac disease is a widely prevalent enteropathy caused by intolerance to gliadin, one of the gluten proteins. We developed two methods for the analysis of gliadin levels. Both methods use flow cytometry and rat antibodies against a 16-residue peptide of gliadin. The peptide is common to the alpha-, beta-, gamma-, and omega-gliadins. METHODS: In the one-site assay, the antigen (gliadin standard or food extract) was adsorbed on 3-mum latex particles. Sensitized particles were then incubated, in this order, with rat anti-gliadin peptide antibodies and anti-rat immunoglobulin G antibodies labeled with fluorescein isothiocyanate. In the two-site assay, the antigen was trapped on the latex particles by rat anti-gliadin antibodies and then measured by the same antibodies labeled with fluorescein. RESULTS: Detection limits were 1 ng/ml for the one-site assay and 10 pg/ml for the two-site assay. The two-site assay displayed gliadin at concentrations above the limit proposed by the Codex Alimentarius in 2 of 40 gluten-free products. CONCLUSION: There is a growing concern that gliadin, even when present in gluten-free foods within the limit fixed by the Codex Alimentarius, over the long term may become toxic to patients with celiac disease. The techniques described in this study provide an opportunity to further decrease the acceptable limit of gliadin in gluten-free foods.
Authors:
Rosanna Capparelli; Ilaria Ventimiglia; Luigi Longobardo; Domenico Iannelli
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Publication Detail:
Type:  Journal Article; Validation Studies    
Journal Detail:
Title:  Cytometry. Part A : the journal of the International Society for Analytical Cytology     Volume:  63     ISSN:  1552-4922     ISO Abbreviation:  Cytometry A     Publication Date:  2005 Feb 
Date Detail:
Created Date:  2005-02-01     Completed Date:  2005-06-02     Revised Date:  2007-07-24    
Medline Journal Info:
Nlm Unique ID:  101235694     Medline TA:  Cytometry A     Country:  United States    
Other Details:
Languages:  eng     Pagination:  108-13     Citation Subset:  IM    
Copyright Information:
(c) 2005 Wiley-Liss, Inc.
Affiliation:
School of Biotechnological Sciences, University of Naples Federico II, Via Università 133, 80055 Portici, Italy. capparel@unina.it
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibody Specificity
Flow Cytometry / methods*,  standards
Gliadin / chemistry*,  immunology
Goats
Microchemistry / methods*,  standards
Nanotechnology / methods*,  standards
Peptide Fragments / chemistry,  immunology
Rats
Reference Standards
Chemical
Reg. No./Substance:
0/Peptide Fragments; 9007-90-3/Gliadin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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