Expression of recombinant parvalbumin from wolf-herring fish and determination of its IgE-binding capability. Issue 4 (4th July 2017)

• Record Type: Journal Article
• Authors: Mohammadi, Mohsen; Mokhtarian, Kobra; Kardar, Gholam Ali; Farrokhi, Shokrollah; Sadroddiny, Esmaeil; Khorramizadeh, Mohammad Reza; Falak, Reza
• Is Part Of: Food and agricultural immunology. Volume 28:Issue 4(2017)
• Page Start: 573
• Page End: 585
• Publication Date: 2017-07-04
• Subjects: Parvalbumin -- wolf-herring fish -- food hypersensitivity -- recombinant protein -- disk ELISA -- Western blotting; Immunochemistry -- Periodicals; Food -- Analysis -- Periodicals; Immunoassay -- Periodicals; 616.079
• DOI: 10.1080/09540105.2017.1306493
• Languages: English
• ISSNs: 0954-0105

Abstract

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.