In vitro assessment of allergenicity features and localization of probable IgE binding regions. (October 2015)

• Record Type: Journal Article
• Authors: Mishra, Ankita; Gaur, S.N.; Lavasa, Shakuntala; Arora, Naveen
• Is Part Of: Food and chemical toxicology. Volume 84(2015)
• Page Start: 181
• Page End: 187
• Publication Date: 2015-10
• Subjects: B-cell epitopes -- Food allergen -- Rice (Oryza sativa) -- Chitinase -- Homology modeling -- in silico tools; Toxicology -- Periodicals; Food poisoning -- Periodicals; Food Poisoning -- Periodicals; Toxicology -- Periodicals; Toxicologie -- Périodiques; Intoxications alimentaires -- Périodiques; Food poisoning; Toxicology; Periodicals; Electronic journals; 615.9
• DOI: 10.1016/j.fct.2015.08.018
• Languages: English
• ISSNs: 0278-6915

Abstract

Abstract: Rice is cultivated as a staple grain crop in many countries, especially in Asia. In the present study, recombinant rice chitinase was expressed, purified and characterized by in silico and immunobiochemical methods. Rice chitinase was affinity purified and it resolved at 24 kDa on SDS-PAGE. Purified protein was analyzed for pepsin resistance, heat stability, and IgE binding using atopic patients' sera. Chitinase was resistant to pepsin digestion and heat treatment at 90 °C for 1 h. It showed significant IgE binding with 7 of 110 patients' sera positive to different food allergens. Homology modeled 3D structure of rice chitinase was used for B cell epitope prediction. In silico predicted B cell peptides were assessed for IgE binding by ELISA using food allergic patients' sera, epitope RC2 showed IgE binding comparable to chitinase. In conclusion, chitinase was identified as a potential allergen and may share cross reactive epitopes with food allergens. Highlights: Expression and purification of rice chitinase protein for immunobiochemical studies. 3D structure of rice chitinase was generated by homology modeling. In silico identification of B-cell epitopes of rice chitinase. Predicted B-cell epitopes were validated by in vitro IgE binding studies. Rice chitinase identified as a potential allergen by in silico and in vitro studies.