Expression of recombinant parvalbumin from wolf-herring fish and determination of its IgE-binding capability |
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| Authors: | Mohsen Mohammadi Kobra Mokhtarian Shokrollah Farrokhi Esmaeil Sadroddiny Mohammad Reza Khorramizadeh |
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| Institution: | 1. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran;2. The Persian Gulf Marine Biotechnology Medicine Research Center, Bushehr University of Medical Sciences, Bushehr, Islamic Republic of Iran;3. Immunology Research Center, Iran University of Medical Sciences, Tehran, Islamic Republic of Iran;4. The Persian Gulf Marine Biotechnology Medicine Research Center, Bushehr University of Medical Sciences, Bushehr, Islamic Republic of Iran;5. Department of Allergy and Immunology, Bushehr University of Medical Sciences, Bushehr, Islamic Republic of Iran;6. Bio-sensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, Tehran, Islamic Republic of Iran |
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| Abstract: | In this study, we produced the recombinant form of parvalbumin from wolf-herring fish and determined its IgE reactivity. Parvalbumin cDNA was sub-cloned into pET28 and expressed in Escherichia coli BL-21. The immunoreactivities of the recombinant and native parvalbumins were compared, and the effect of calcium binding was determined by sera from 25 fish-allergic patients. ELISA and Western blotting confirmed similar IgE-reactivities of the recombinant and native proteins and confirmed that this phenomenon is highly dependent on calcium binding. The recombinant protein was 94.5% similar to carp parvalbumin (Cyp c1). Approximately 72% of patients reacted strongly with recombinant parvalbumin, 80% of them reacted with the native form and only 56% showed IgE reactivity with crude extract. Because the IgE-binding capacity of recombinant wolf-herring parvalbumin is retained and is highly similar to Cyp c1, the wild and hypoallergenic forms of this allergen could be used for diagnosis and immunotherapy of fish allergy, respectively. |
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| Keywords: | Parvalbumin wolf-herring fish food hypersensitivity recombinant protein disk ELISA Western blotting |
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