Identification of the major allergenic epitopes of Eriocheir sinensis roe hemocyanin: A novel tool for food allergy diagnoses |
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| Institution: | 1. School of Medical Laboratory, Tianjin Medical University, 1 Guangdong Road, Hexi District, Tianjin 300203, China;2. Academy of Traditional Chinese Medicine Affiliated Hospital, 354 Beima Road, Hongqiao District, Tianjin, China;3. Department of Medical Laboratory, Tianjin Children’s Hospital, 225 Machang Road, Hexi District, Tianjin, China;1. Laboratório de Nanobiotecnologia, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Amazonas, Bloco 2E, sala 248, Uberlândia 38400-902, Minas Gerais, Brazil;2. Laboratório de Biomateriais, Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia, Av. Amazonas, Bloco 2E, Uberlândia 38400-902, Minas Gerais, Brazil;3. Laboratório de Filmes Poliméricos e Nanotecnologia, Instituto de Química, Universidade Federal de Uberlândia, Av. João Naves de Ávila 2121, Uberlândia 38408-100, Minas Gerais, Brazil;4. Unidade de Investigação em Reumatologia, Instituto de Medicina Molecular, Av. Prof. Egas Moniz, Lisboa 1649-028, Portugal;5. Unidade de Reumatologia Pediátrica, Hospital de Clínicas, Faculdade de Medicina da Universidade Federal de Uberlândia, Av. Pará 1720, Bloco 2U, Uberlândia 38400-902, Brazil;6. Department of Medical Microbiology and Immunology, University of California Davis, 1 Shields Avenue, Tupper Hall, Rm. 3146, Davis, CA 95616, USA;7. Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Av. Prof. Egas Moniz, Lisboa 1649-028, Portugal;1. Pediatric Urology Research Laboratory, Nemours Biomedical Research/Alfred I. duPont Hospital for Children, Wilmington, Delaware;2. Tissue Engineering and Regenerative Medicine Research Laboratory, Nemours Biomedical Research/Alfred I. duPont Hospital for Children, Wilmington, Delaware;3. Department of Mechanical Engineering, University of Delaware, Newark, Delaware;4. Department of Biomedical Engineering, University of Delaware, Newark, Delaware;1. Department of Anatomy, Brain Research Institute, Chungnam National University School of Medicine, Munwha-ro 266, Jung-gu, Daejeon, 301-747, Republic of Korea;2. Department of Pediatrics, Chungnam National University Hospital, Brain Research Institute, Chungnam National University School of Medicine, Munwha-ro 282, Jung-gu, Daejeon, 301-721, Republic of Korea;3. Department of Physiology, Brain Research Institute, Chungnam National University School of Medicine, Munwha-ro 266, Jung-gu, Daejeon, 301-747, Republic of Korea;4. Department of Plastic Surgery, Chungnam National University Hospital, Munwha-ro 282, Jung-gu, Daejeon, 301-721, Republic of Korea;5. Anesthesia and Pain Medicine, Chungnam National University Hospital, Munwha-ro 282, Jung-gu, Daejeon, 301-721, Republic of Korea;6. Department of Chemical and Biological Engineering, The University of Alabama, Tuscaloosa, AL 35487, USA;1. College of Food and Biological Engineering, Fujian Collaborative Innovation Center for Exploitation and Utilization of Marine Biological Resources, Jimei University, 43 Yindou Road, Xiamen 361021, Fujian, China;2. School of Biosciences, Sutton Bonington Campus, Loughborough LE12 5RD, UK;1. School of Food and Chemical Engineering, Beijing Technology and Business University, Beijing 100048, China;2. Institute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Beijing 100850, China;3. Department of Pharmacy, Dalian Municipal Friendship Hospital, Dalian 116100, China;4. Department of Pediatrics, Children׳s Memorial Research Center, Northwestern University Feinberg School of Medicine, Chicago, IL, USA |
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| 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. |
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| Keywords: | Food allergy Hemocyanin Linear epitope Recombinant allergen |
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