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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
- Other title
- Type of item
- Degree grantor
University of Alberta
- Author or creator
- 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)
Faculty of Pharmacy and Pharmaceutical Sciences
- Date accepted
- Graduation date
Doctor of Philosophy
- Degree level
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.
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- 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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