Production Of Monoclonal Antibodys Of The Main Allergen In River Shrimp And Analysis Of The Common Epitopes Between The Different Allergens

AIM:To identify the allergens in River shrimp, and to isolate, purify and analyze the main allergen components. Preparing the monoclonal antibody of the main shrimp allergen to analyze the shared epitope of the different allergens in river shrimp and the cross reaction between different shrimps and crabs, which will lay a foundation for shrimp allergy clinical diagnosis and shrimp allergen vaccine design.METHOD:Three main River shrimp allergens were purified by ammonium sulfate precipitation, Sephadex G-50 chromatography and DEAE-exchange chromatography from the total river shrimp proteins. The BALB/C mice were immunized with purified 36kD allergen to produce its monoclonal antibody. Indirect ELISA, indirect competitive ELISA and Western Blot were exploited to analyze the common epitopes of the river shrimp allergens and cross reaction between river shrimp, sea shrimp and crab.RESULTS:The purified 21kD,36kD,80kD allergens were estimated to be about 97.5%,94.2% and 85.3% pure. Western blot demonstrated that the reaction rate between main allergens 21kD, 36kD,80kD and shrimp allergic patients'sera IgE were 43.3%,76.7%,36.7%.1D11,2C4, 3C4,4A3,4E3,4A5B11,4A5C10 and 4A5D5 McAbs were obtained, and the title of 1D11,3C4 and 4E3 wasl:64000,1:128 000,1:6400. The results of indirect ELISA, competitive indirect ELISA and western blot showed:the 1D11,3C4 and 4E3 can recognized the eptiope of shrimp allergens; 1D11,2C4,4A5B1 land 4A5D5 can react with the other allegens in river shrimp. The indirect competitive ELISA demonstrated that the allergens in river shrimp, sea shrimp and crab can obviously surpress each other.CONSULUTION:The 21kD,36kD and 80kD proteins are the important allergens in river shrimp, and the 36kD protein is the primary main allergen, with the highest allergenicity and sensitization rate. There are some common epitopes in 21kD,36kD and 80kD allergen component. Cross reaction were happened between river shrimp, sea shrimp and crab; 36kD allergen component share common epitopes with 36kD allergen component in sea shrimp and 23kD,36kD,70kD protein components in crab.