title: Identification of the major allergenic epitopes of Eriocheir sinensis roe hemocyanin: A novel tool for food allergy diagnoses.
abstract: 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.
authors.affiliation: School of Medical Laboratory, Tianjin Medical University, 1 Guangdong Road, Hexi District, Tianjin 300203, China.
authors.forename: Yingying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: School of Medical Laboratory, Tianjin Medical University, 1 Guangdong Road, Hexi District, Tianjin 300203, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Academy of Traditional Chinese Medicine Affiliated Hospital, 354 Beima Road, Hongqiao District, Tianjin, China.
authors.forename: Shaoshen
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Medical Laboratory, Tianjin Children's Hospital, 225 Machang Road, Hexi District, Tianjin, China.
authors.forename: Jiayi
authors.initials: J
authors.lastname: Zhang
authors.affiliation: School of Medical Laboratory, Tianjin Medical University, 1 Guangdong Road, Hexi District, Tianjin 300203, China.
authors.forename: Tiantian
authors.initials: T
authors.lastname: She
authors.affiliation: School of Medical Laboratory, Tianjin Medical University, 1 Guangdong Road, Hexi District, Tianjin 300203, China.
authors.forename: Juanjuan
authors.initials: J
authors.lastname: Yan
authors.affiliation: School of Medical Laboratory, Tianjin Medical University, 1 Guangdong Road, Hexi District, Tianjin 300203, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Bian
authors.affiliation: School of Medical Laboratory, Tianjin Medical University, 1 Guangdong Road, Hexi District, Tianjin 300203, China. Electronic address: lhq@tmu.edu.cn.
authors.forename: Huiqiang
authors.initials: H
authors.lastname: Li
mesh_terms: D000328:Adult; D000485:Allergens / Q000276:immunology; D000818:Animals; D003386:Brachyura / Q000276:immunology*; D002675:Child, Preschool; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005512:Food Hypersensitivity / Q000175:diagnosis* / Q000276:immunology*; D006433:Hemocyanins / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D016056:Immunodominant Epitopes / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology; D007223:Infant; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology; D055815:Young Adult
chemical_list: D000485:Allergens; D016056:Immunodominant Epitopes; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D006433:Hemocyanins
keywords: Eriocheir sinensis; Food allergy; Hemocyanin; Linear epitope; Recombinant allergen