Recognition of an Extensive Range of IgE-reactive Proteins in Cod Extract

Allergy. 1998 Jan;53(1):42-50. doi: 10.1111/j.1398-9995.1998.tb03772.x.

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 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Allergens / analysis
  • Allergens / immunology*
  • Allergens / isolation & purification
  • Animals
  • Antibodies, Monoclonal / immunology
  • Blotting, Western / methods
  • Chelating Agents / pharmacology
  • Child
  • Child, Preschool
  • Chromatography, DEAE-Cellulose
  • Edetic Acid / pharmacology
  • Electrophoresis, Polyacrylamide Gel
  • Epitopes / immunology
  • Fishes / immunology*
  • Food Handling / methods
  • Food Hypersensitivity / immunology*
  • Humans
  • Immunoglobulin E / immunology*
  • Infant
  • Middle Aged
  • Parvalbumins / immunology
  • Proteins / analysis
  • Proteins / immunology*
  • Proteins / isolation & purification*
  • Radioimmunoassay

Substances

  • Allergens
  • Antibodies, Monoclonal
  • Chelating Agents
  • Epitopes
  • Parvalbumins
  • Proteins
  • Immunoglobulin E
  • Edetic Acid