Purification And Identification Of Immunoactivity Of Allergen Protein From Several Familiar Food Allergens

Objective: To develop ana optimize the assay of detect specific IgE anddirectionally releasing histamines through copying the food allergic models, so as to provide the multiple allergens detection technique in vitro , which was specific, sensitive and safe, would be used to substitute for the prick test in clinic, and pave the ways to express the recombinant allergens and standardize the allergens.Methods: Food allergen extract in milk, crab and turtle were produced, and thefood allergic animal models were established with them. The specific IgE and IgG of food allergens in sera of the model mice were measured by indirect ELISA. The peritoneal mast cells were collected, stimulated by the allergen, and the released contents of histamine were detected by fluorescence assay. The allergic components of the food allergen were analized by western-blotting, and isolated and purified by saturated sulphate ammonium.Results: Allergic BALB/c models of whey protein and crab protein had beenestablished by hypodermic injected with the adjuvant of aluminum hydroxide. The levels of specific IgE in allergic mice models were remarkably higher than that in control ones, which indicated the success of developing allergic animal models. The optimized method of immune was established by hypodermic injection with the adjuvant of aluminum hydroxide. The base value of the normal bred group was much higher than that of the BALB/c mice bred for two generations on a milk-free diet. The results of release histamine ratio at 14th day after stimulation were well accordance with the results of specific IgE.Conclusion: The detection of specific IgE and the histamine releasing ratio of theirperitoneal mast cells might become a new method to characterize and assess food allergens, which were significant for the studies of allergy diagnosis , treatment, and could be used to detect the allergens in gene modified foods.