The Research On Technology Of Elimination And Detection To Major Cow's Milk Allergens

In this paper, using trypsin, papain and bromelain hydrolyze milk, through orthogonal experiment and using the serum of the person which is sensitive to milk allergen to optimize the best method of hydrolyze milk allergy. Research shows that the optimal reaction conditions for trypsin: temperature at 40°C, pH8.0, the rate of quality between enzyme and substrate is 5% ,hydrolysis time is 3 hours;the best conditions for the hydrolysis of papain: 65°C, pH6.5, the rate of quality between enzyme and substrate is 4%, hydrolysis time is 3 hours; the best conditions for the hydrolysis of bromelain: 60°C, pH6.5, the rate of quality between enzyme and substrate is 4%, hydrolysis time is 2 hours. In the best enzymatic hydrolysis conditions , hydrolyze the milk by add the enzyme in turn and after hydrolysis of the milk, the result by ELISA test between cow's milk allergy and serum specific IgE of the person is in the least and it has almost no demonstrated antigen from the milk. Analysis shows that several of the major allergens in milk have epitopes which are the enzyme that use in the experiment recognition sites . Enzyme's molecules attack the site where in the spaces of the epitope on the protein and damage the structure of the milk allergens, thus weakening or eliminating the allergen of milk proteins.The experimental animal model established aim to obtain allergic serum as contrast .The experimental animal model serum be used in the test of the serum's IgE specificity . The results indicate that the serum's IgE from milk allergy patient's is specific allergy to cow's milk protein and established the foundation with specificity detection of this experiment. And we established the method by use of cow's milk and sensitively serum to detect the degree of enzyme hydrolysis of the milk allergen protein. By the use of animal model, we found adjuvant, IFN and IL-2 can improve the quantity of the special IgE and IgG, and the IL-11 can decrease the quantity of the special IgE and IgG.