Expression of Recombinant Parvalbumin From Wolf-Herring Fish and Determination of Its Ige-Binding Capability Publisher



Mohammadi M^{1, 2} ; Mokhtarian K³ ; Kardar GA⁴ ; Farrokhi S^{2, 5} ; Sadroddiny E¹ ; Khorramizadeh MR^{1, 6} ; Falak R^{3, 7} Show Affiliations
Source: Food and Agricultural Immunology Published:2017

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. © 2017 Informa UK Limited, trading as Taylor & Francis Group.