Document Detail

Multiple protein extract microarray for profiling human food-specific immunoglobulins A, M, G and E.
MedLine Citation:
PMID:  20974146     Owner:  NLM     Status:  MEDLINE    
Existing food immunoglobulin (Ig) tests require large volumes of serum, are limited to one immunoglobulin class, are not amenable to high throughput analysis and only give a limited picture of the immunological response to food antigens. Conversely a new generation of Component Resolved Diagnostic systems using pure proteins is highly specific and totally dependent on the availability of the protein in its recombinant or natural origin form. Here we demonstrate a proof-of-concept of a microarray test based on protein extracts of food components. Our approach relies on innovations on three different fronts: the novelty of using arrayed food samples sequentially extracted with detergent and chaotropic agents, the ability to measure four different Ig classes simultaneously and the ability to analyse the generated data via a suitable bioinformatics/statistical analysis interface. This approach combines high numerical power of microarrays with automation, high throughput analysis and enables detailed investigation of the Ig profiles to food antigens. The prototype shown contains extracts of approximately 350 food ingredients that cover most of the food products found in the UK. Here we showed that the use of a sequential extraction technique to solubilise and then denature food samples has its benefits in the assessment of variations in antigenicity when tested with human sera. A patient dependent degree of class specificity was observed with human sera (IgG specificity correlates well with IgA>IgM>>>>>IgE). Besides generating a simultaneous profile for IgA, IgM, IgG and IgE the array system has shown good discrimination between challenge responders in atopic and non-atopic individuals. Poly- and mono-specific IgE responders were easily identified. The mathematical modelling of specific IgE content showed good correlations when compared with established IgE antibody testing assay (UniCAP). Although in its proof-of-principle stages, the immune profiling technique described here has the potential to provide unique insights into exposure/sensitization and establish relationships between specific immunoglobulin classes and subclasses against food protein antigens. In further developments, the immune profiling technique could also be extended to other related areas such as parasite and bacterial gut infection. Full analyses of large longitudinal and retrospective clinical trials are on going to determine the positive and negative predictive values of the technique.
N K Renault; S R Gaddipati; F Wulfert; F H Falcone; L Mirotti; P J Tighe; V Wright; M J C Alcocer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-10-23
Journal Detail:
Title:  Journal of immunological methods     Volume:  364     ISSN:  1872-7905     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  2011 Feb 
Date Detail:
Created Date:  2011-01-10     Completed Date:  2011-05-17     Revised Date:  2014-02-20    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  21-32     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier B.V. All rights reserved.
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MeSH Terms
Allergens / immunology,  metabolism*
Automatic Data Processing
Cell Extracts
Computational Biology
Food Hypersensitivity / blood,  diagnosis*,  immunology
Great Britain
High-Throughput Screening Assays
Immunoglobulins / blood*
Models, Theoretical
Predictive Value of Tests
Protein Array Analysis / methods*
Proteins / immunology,  metabolism*
Sensitivity and Specificity
Grant Support
BB/FOF/268//Biotechnology and Biological Sciences Research Council; G0502250//Medical Research Council
Reg. No./Substance:
0/Allergens; 0/Cell Extracts; 0/Immunoglobulins; 0/Proteins

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