Antigenicity Of Common Carp Parvalbumin

Fish is a good resource of food protein for people. But some people suffer from allergic reaction against fish protein, their heath are seriously threatened. Studies have shown that the major food allergen of fish such as common carp and cod is parvalbumin. So the study of carp parvalbumin will be useful to the prevention of food allergy and the production of low-allergenic products.The investigation is composed of three parts including the purification of common carp parvalbumin, the preparation of rabbit anti-carp parvalbumin polyclonal antibody and the immune cross reaction between common carp parvalbumin and crude extraction of black carp, grass carp and silver carp.Common carp parvalbumin was purified using ion-exchange chromatography on DEAE-Sepharose Fast Flow gel with loading sample extracted in buffered saline. SDS-PAGE and PAGE were used to identify the purity of the protein. The result of SDS-PAGE and PAGE showed that high purified common carp parvalbumin can be obtained. The purity of common carp parvalbumin was more than 90%.An indirect ELISA was developed to detect the titre of the specific rabbit serum and a protocol to prepare specific polyclonal antibody against common carp parvalbumin was established. Two Japanese male rabbits were immunized with purified common carp parvalbumin and the titre of the serum of the two rabbits was 1:560,000 and 1:720,000 respectively by indirect ELISA. The assessment of common carp parvalbumin's antigenicity was performed on the evaluation of immunogenicity and immunoreactivity by injecting common carp parvalbumin into the rabbits and using indirect competitive ELISA. The purified common carp parvalbumin's antigenicity was preserved well.Through the Western blotting and indirect competitive ELISA tested, the immune cross reaction between common carp parvalbumin and crude extraction of black carp, grass carp and silver carp was validated, and the primary protein molecular weight which has an immune reaction was about 12 kDa.