Recognition of an extensive range of IgE-reactive proteins in cod extract.
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 post-rigor 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 IgE-reactive 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 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.
Dory
D
D
Laboratoire de Pathologie Cellulaire et Moléculaire en Nutrition, EP CNRS 616, Faculté de Médecine de Nancy, Vandoeuvre, France.
Chopin
C
C
Aimone-Gastin
I
I
Gueant
J L
JL
Guerin
L
L
Sainte-Laudy
J
J
Moneret-Vautrin
D A
DA
Fleurence
J
J
D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western / Q000379:methods; D002614:Chelating Agents / Q000494:pharmacology; D002648:Child; D002675:Child, Preschool; D002848:Chromatography, DEAE-Cellulose; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D005399:Fishes / Q000276:immunology*; D005511:Food Handling / Q000379:methods; D005512:Food Hypersensitivity / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D007223:Infant; D008875:Middle Aged; D010320:Parvalbumins / Q000276:immunology; D011506:Proteins / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification*; D011863:Radioimmunoassay
D000485:Allergens; D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D000939:Epitopes; D010320:Parvalbumins; D011506:Proteins; D007073:Immunoglobulin E; D004492:Edetic Acid