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Monoclonal Antibodies to Recombinant Fag e 3 Buckwheat Allergen and Development of a Two-site ELISA for Its Quantification

 Kyoung Yong Jeong  ;  Kyung Hee Park  ;  Jae-Hyun Lee  ;  Jung-Won Park 
 Allergy Asthma & Immunology Research, Vol.9(5) : 417-422, 2017 
Journal Title
 Allergy Asthma & Immunology Research 
Issue Date
2-site ELISA ; Buckwheat ; Fag e 3 ; vicilin
PURPOSE: Buckwheat is a major cause of anaphylaxis, and Fag e 3 is the key major allergen in buckwheat. However, an immunoassay system for the quantification of Fag e 3 has yet to be developed. METHODS: We developed a 2-site enzyme-linked immunosorbent assay (ELISA) using monoclonal antibodies (mAbs) produced against recombinant Fag e 3. We applied this ELISA to quantify native Fag e 3 in total buckwheat extract. RESULTS: Four clones of mAbs were produced, and all recognized vicilin allergens not only from buckwheat, but also from peanut and walnut. However, the ELISA using these antibodies was only able to quantify Fag e 3 in the total extract after addition of 1% sodium dodecyl sulphate (SDS) and heating, which facilitated dissociation of the allergen. The detection limit of the developed 2-site ELISA was 0.8 μg/mL. The measurement of Fag e 3 in the total extract of buckwheat showed that approximately 12% of protein in total buckwheat extract was Fag e 3. CONCLUSIONS: We have developed an ELISA system for the quantification of the group 3 buckwheat allergen, Fag e 3, specifically. This assay will be useful for standardization of buckwheat allergens and monitoring of buckwheat contamination in foods.
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1. College of Medicine (의과대학) > Dept. of Internal Medicine (내과학교실) > 1. Journal Papers
5. Research Institutes (연구소) > Institute of Allergy (알레르기연구소) > 1. Journal Papers
Yonsei Authors
박경희(Park, Kyung Hee) ORCID logo https://orcid.org/0000-0003-3605-5364
박중원(Park, Jung Won) ORCID logo https://orcid.org/0000-0003-0249-8749
이재현(Lee, Jae Hyun) ORCID logo https://orcid.org/0000-0002-0760-0071
정경용(Jeong, Kyoung Yong) ORCID logo https://orcid.org/0000-0001-9887-1426
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