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

  • Author / Creator
    Gujral, Naiyana
  • 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.

  • Subjects / Keywords
  • Graduation date
    2013-11
  • Type of Item
    Thesis
  • Degree
    Doctor of Philosophy
  • DOI
    https://doi.org/10.7939/R31R6NB49
  • License
    This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.
  • Language
    English
  • Institution
    University of Alberta
  • Degree level
    Doctoral
  • Department
    • Faculty of Pharmacy and Pharmaceutical Sciences
  • Specialization
    • Pharmaceutical Sciences
  • Supervisor / co-supervisor and their department(s)
    • Lobenberg, Raimar (Pharmaceutical Sciences)
    • Sunwoo, Hoon (Pharmaceutical Sciences)
  • Examining committee members and their departments
    • Miyoung, Suh (Human Nutritional Sciences)
    • Sharon Marsh (Pharmaceutical Sciences)
    • Thomson, Alan (Medical Sciences)
    • Doschak, Michael (Pharmaceutical Sciences)