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Permanent link (DOI): https://doi.org/10.7939/R31R6NB49

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Antibody-based Diagnostic and Therapeutic Approaches on Gluten-sensitive Enteropathy Open Access

Descriptions

Other title
Subject/Keyword
Gluten-sensitive enteropathy
Celiac disease
Gluten
Oral antibody
IgY antibody
Diagnostics
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Gujral, Naiyana
Supervisor and department
Sunwoo, Hoon (Pharmaceutical Sciences)
Lobenberg, Raimar (Pharmaceutical Sciences)
Examining committee member and department
Miyoung, Suh (Human Nutritional Sciences)
Sharon Marsh (Pharmaceutical Sciences)
Thomson, Alan (Medical Sciences)
Doschak, Michael (Pharmaceutical Sciences)
Department
Faculty of Pharmacy and Pharmaceutical Sciences
Specialization
Pharmaceutical Sciences
Date accepted
2013-08-27T14:44:25Z
Graduation date
2013-11
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Gluten-sensitive enteropathy, called Celiac disease (CD), is one of the most frequent autoimmune diseases, occurring in 1% people worldwide, upon gliadin ingestion. Currently, the only treatment available for CD individual is a strict life-long gluten-free diet. Chicken egg yolk immunoglobulin Y (IgY) is produced and examined for its efficacy in vitro, ex vivo, and in vivo to prevent enteric absorption of gliadin. This antibody was also used to develop sensitive and rapid detection kits for gluten. The extracted toxic gliadin was immunized into chickens inducing humoral immune response to produced gliadin-specific IgY antibodies. The IgY antibody was separated from non-protein component in egg yolk and was purified by gel chromatography, showing > 95% purity on electrophoresis. One gram of purified IgY antibody contains 79 mg of specific anti-gliadin IgY antibody based on quantitative ELISA technique. Under in vitro simulated gastric and intestinal conditions, competitive ELISA demonstrated that 1.5 mg anti-gliadin IgY completely neutralized 6.6 g gliadin in test tube. Using Caco2 cell culture as ex vivo test, anti-gliadin IgY prevented gliadin absorption (at a ratio of 1:3,000), resulting in no pro-inflammatory response (TNF-α and IL-1β). In-vivo BALB/c mice study showed that 31 μg specific anti-gliadin IgY antibody prevented 100 mg gliadin absorption in the gastrointestinal tract when both antibody and gliadin were orally fed. The developed antibody was used to also develop sensitive double antibody sandwich ELISA (DAS-ELISA) Immunoswab and Immunostrip assay based on anti-gliadin IgY and biotinylated monoclonal antibody (mAb) showing a detection limit of 4 ng/mL, 1.25 µg/mL and 0.25 µg/mL, respectively. Anti-gliadin IgY has potential to be used as an oral passive antibody therapy to treat CD. This CD therapeutic candidate may provide an effective means of preventing CD when co-administered with gliadin contained food. Further clinical studies are warranted to test anti-gliadin IgY formulation in CD subjects exposed to gluten. The combination of anti-gliadin IgY and biotinylated mAb provided reliable, sensitive and inexpensive tools for the detection of gluten in gluten-free and gluten-contained food products.
Language
English
DOI
doi:10.7939/R31R6NB49
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
Gujral N, Freeman HJ, Thomson AB. Celiac disease: prevalence, diagnosis, pathogenesis and treatment. World Journal of Gastroenterology 2012;18(42):6036-6059.Gujral N, Lobenberg R, Suresh MR, Sunwoo HH. In-vitro and in-vivo binding activity of chicken egg yolk immunoglobulin Y (IgY) against gliadin in food matrix. Journal of Agricultural Food Chemistry 2012;60(12):3166-3172.Gujral N, Suresh MR, Sunwoo HH. Quantitative double antibody sandwich ELISA for the determination of gliadin in foods. Journal of Immunoassay and Immunochemistry 2012; 33:339–351.Sunwoo H, Gujral N, Lutz S, Suresh MR. Double antibody sandwich ELISA and rapid Immunoswab assay for detection of gliadin in food. Food and Agriculture Immunology 2012;23(2): 169-181.

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