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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Healthy Processed Foods Research » Research » Publications at this Location » Publication #398369

Research Project: Prevention of Obesity Related Metabolic Diseases by Bioactive Components of Food Processing Waste Byproducts and Mitigation of Food Allergies

Location: Healthy Processed Foods Research

Title: IgE binding epitope mapping with TL1A tagged peptides

item Zhang, Yuzhu
item Bhardwaj, Shilpa
item Vilches, Ana
item Breksa, Andrew
item LYU, SHU-CHEN - Stanford University School Of Medicine
item CHINTHRAJAH, SHARON - Stanford University School Of Medicine
item NADEAU, KARI - Stanford University School Of Medicine
item JIN, TENGCHUAN - University Of Science And Technology Of China

Submitted to: Molecular Immunology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 12/1/2022
Publication Date: 12/15/2022
Citation: Zhang, Y., Bhardwaj, S.R., Vilches, A.M., Breksa III, A.P., Lyu, S., Chinthrajah, S., Nadeau, K., Jin, T. 2022. IgE binding epitope mapping with TL1A tagged peptides. Molecular Immunology. 153:194-199.

Interpretive Summary: Food allergies have become a significant health concern for patients and their families, the food industries, and the public. A monumental aspect of food allergies is the association of allergens with immunoglobulin E (IgE). The part of an allergen to which an IgE binds is called an IgE binding epitope. Information about IgE binding epitopes can be used for accurate and reproducible diagnoses of allergies. However, such information is very limited at present. Mapping linear epitopes of an allergen requires the production of overlapping peptides or small pieces of the allergen. The difficulties in synthesizing and purifying a large number of peptides often impede the identification of IgE epitopes of allergens. Therefore, developing a reliable method for producing peptides with various properties is very important. In this study, an expression system was designed, and its application to IgE epitope mapping was demonstrated by identifying IgE epitopes of the peanut allergen Ara h 2. This new method may enable researchers to obtain information about IgE binding epitopes of other allergens, thus allowing better diagnosis of allergies and understanding of the allergenicity of proteins.

Technical Abstract: Linear IgE epitopes play essential roles in peanut and tree nut allergies. Overlapping peptides spanning the entire length of the protein sequences of food allergens were used in dot blot and microarray experiments, and the dominant linear epitopes of peanut allergens Ara h 1-3 were identified. However, prevalent IgE epitopes for most known food allergens have not been mapped. One of the challenges for linear IgE epitope mapping is the production of peptides. Chemically synthesizing peptides is currently the primary method of peptide production. It is expensive, and the process has inherent challenges in scaling up the production and purification of the synthesized peptides. To overcome this problem, we have constructed a plasmid vector for expressing peptides sandwiched between an N-terminal His-tag and a trimeric protein. The vector was used to make overlapping peptides derived from peanut allergens Ara h 2. All the peptides were successfully expressed and purified, and the method was applied to identify IgE binding epitopes of Ara h 2. This system can also be used in future research to produce agents for component- and epitope-resolved food allergy diagnoses.