Recognition of an extensive range of IgE-reactive proteins in cod extract |
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Authors: | D Dory C Chopin I Aimone-Gastin J L Gueant L Guerin J Sainte-Laudy D A Moneret-Vautrin J Fleurence |
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Institution: | Laboratoire de Pathologie Cellulaire et Moléculaire en Nutrition, EP CNRS 616, Facultéde Médecine de Nancy, Avenue de la Forêt óe Haye, BP 184, 54505 Vandœuvre Cedex;Laboratoire de Qualitéet Physico-Chimie, IFREMER, rue de I'lle d'Yeu, BP 21105, 44311 Nantes Cedex 03;SociétéAllerbio, 8 rue porte aux champs, 55270 Varennes-en-Argonne;CERBA, rue de I'Equerre, 95310 Saint Ouen L'Aumone;Service d'Allergologie et Immunologie Clinique, CHU Nancy, Hôpital Central, 29 avenue du Maréchal De Lattre de Tassigny, 54000 Nancy, France |
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Abstract: | Allergy to fish is one of the most common food allergies. Gad c 1 is the only fish allergen which has been purified and characterized. Other allergens have been detected by Western blot in cod extracts. We have now improved the Western-blot procedure in order to characterize fish IgE-reactive proteins from extracts prepared under different conditions: pre-rigor mortis and postrigor mortis. EDTA addition or not. and DEAE ion-exchange chromatography. Several IgE-reactive protein bands have been identified over a wide molecular-weight range. In particular, the 104- and 130-kDa IgEreactive protein bands were detected. These new bands may correspond to aggregates, as EDTA increased the relative amount of the 60-, 67-, 104-, and 130-kDa IgE-reactive protein bands in Western blot. All these bands were also detected by an antiparvalbumin monoclonal antibody, specific to the first calcium-binding site. The longer period of storage increased the relative amounts of the 41-, 80-, 104-. and 130-kDa IgE-reactive protein bands. The 18-kDa band was detected only in fish stored for several days. In conclusion, we have described IgE-reactive protein bands over a wide molecular-weight range (12–130 kDa) in Western blot of cod extract, and shown that EDTA and storage conditions may influence the relative distribution of IgE-reactive protein bands. |
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Keywords: | calcium-binding site cod IgE-reactive protein bands fish allergy hypersensitivity ion exchange chromatography Western blotting |
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