Rapid detection of fish major allergen parvalbumin by surface plasmon resonance biosensor

Seafood allergy is a common and major cause of food allergy in adults. In recent years, seafood allergy has become a serious problem with the increase of seafood consumption. To develop a rapid allergen detection method based on the affinity of antigen-antibody interaction, fish major allergen, parvalbumin, was used for kinetic analysis by a surface plasmon resonance (SPR) biosensor. Anti-parvalbumin murine monoclonal antibody (MAb) EG8 was immobilized onto a carboxymethyl dextran (CMD) surface. By the injection of various concentrations of purified carp parvalbumin (CPa), a standard curve and the affinity constants (K(D) and k(ass)) for the MAb EG8-CPa model system were determined. In addition, kinetic data were also obtained by the injection of serial dilutions of extracts from seafood products: sardine fish cake (tsumire) and dried skipjack tuna (katsuonut). Sardine tsumire and katsuonut contained 0.11 mg/kg and 0.39 mg/kg parvalbumins, respectively, where affinity constants K(D) and k(ass) were almost similar among parvalbumins from different sources. In the SPR system, the allergen can be detected only for 5 min according to the allergen-MAb binding interaction. Consequently, by the use of a SPR biosensor, kinetic analysis based on the allergen specific MAb would be a rapid and powerful tool for allergen detection and quantification.