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Identification of the major allergenic epitopes of Eriocheir sinensis roe hemocyanin: A novel tool for food allergy diagnoses
- Zhang, Yingying, Zhu, Lina, Li, Shaoshen, Zhang, Jiayi, She, Tiantian, Yan, Juanjuan, Bian, Ying, Li, Huiqiang
- Molecular Immunology 2016 v.74 pp. 125-132
- Eriocheir sinensis, Escherichia coli, allergenicity, allergens, antibodies, bioinformatics, crab meat, crabs, dose response, enzyme-linked immunosorbent assay, food allergies, hemocyanin, immunoblotting, immunodominant epitopes, immunoglobulin E, patients, polyacrylamide gel electrophoresis, prediction, recombinant fusion proteins, China
- Crab meat and roe are highly nutritious delicacies in China. While extensive research has been conducted for allergens derived from crab-meat, data relevant to the allergenic potential of crab roe derived proteins, of which hemocyanin is a principal contender, are almost entirely absent. Using bioinformatics prediction and IgE-binding assays, the three principal immunodominant epitopes of hemocyanin were identified and then combined as a single recombinant fusion protein (rHc). This together with the full-length recombinant protein (Hc) were expressed in Escherichia coli and subsequently identified by SDS-PAGE and immunoblotting. Ninety-five percent of our patients were found to carry rHc-specific IgE antibodies by ELISA. Dot-blot inhibition, together with ELISA inhibition studies, showed that pre-incubation of patient sera with the recombinant epitope protein could inhibit26% to 63% (mean: 50%) of IgE binding to immobilized, full-length Hc and the dose-response curve represents as a sigmoid shape. The recombinant protein (rHc) represents a versatile biologic tool with which to diagnose and investigate therapies for E. sinensis allergy.