Identification,isolation,purification of allergens in shrimp and allergic analysis for the main allergens

AIM:To identify the allergens in shrimp,and to isolate,purify and analyze the main allergen components.METHODS:The total shrimp proteins were extracted by PBS,the allergens were identified with 11 shrimp allergic patients’ serum IgE by Western blot.Three main shrimp allergens 21 000,36 000 and 80 000 were purified by ammonium sulfate precipitation,Sephadex G-50 chromatography and DEAE-exchange chromatography.Western blot and indirect ELISA were used to confirm and analyze allergenicity of the three main allergens.RESULTS:Western blot demonstrated that there were nine allergen components in shrimp proteins,and IgE binding to 21 000,36 000 and 80 000 shrimp allergen by 5,7,5 (36.4%,63.6%,45.5%) of 11 shrimp allergic patients' sera.Results of indirect ELISA showed that the binding absorbency of allergic patients’ sera IgE with the three main purified allergens were all higher than that with shrimp protein extraction.CONCLUSION:There are at least 9 allergens in shrimp;the 21 000,36 000 and 80 000 proteins are the main allergen components;and the 36 000 protein is the primary main allergen,with the highest allergenicity and sensitization rate.In further investigation,we will use monoclonal antibody technique to verify weather the 21 000,36 000,80 000 allergens have common epitope,which will lay a foundation for shrimp allergy clinical diagnosis and shrimp allergen vaccine design.