Development of a monoclonal antibody-based competitive enzyme linked-immunosorbent assay (c-ELISA) for quantification of silver carp parvalbumin

Parvalbumin (PV) is a major allergen in fish. A monoclonal antibody (B2-E1) against silver carp parvalbumin was prepared in the present study. Western blot analysis indicated that the prepared monoclonal antibody was specific to PV in fish muscle extract. A competitive enzyme-linked immunosorbent assay (c-ELISA) was then developed to quantify the amount of PV in silver carp using B2-E1. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed c-ELISA were 0.04 and 0.3 mg/kg food, respectively. The c-ELISA was specific for detecting PV from fish, and the intra- and inter-assay coefficients of variation were 4.4% and 8.4%, respectively. The c-ELISA method was further applied to quantify PV from various fish species, and demonstrated that the PV content was 4-20 times higher in the white muscle than that in the dark muscle, which was coincident with the previous reports. Moreover, c-ELISA was used to trace the PV during fish surimi (fish ball) production, and the results indicated that washing process could effectively but not completely remove PV from fish ball. Our present study indicated that this c-ELISA method may be applied to monitor trace amounts of fish allergen PV during fishery product processing.

► An anti-silver carp parvalbumin (PV) mAb with high specificity was prepared. ► A specific c-ELISA method was developed to monitor silver carp PV in foods. ► The established c-ELISA was highly reproducible and sensitive. ► Washing process could effectively but not completely remove PV from fish balls.