Bioinformatics Analtsis And Establishment Of Detection Method For Sesame Allergens

Food allergy disease is one of the major global public health problems,and the number of people affected by food allergy disease is on the rise.Therefore,research on sesame allergens has become particularly important,such as predicting the structure of sesame allergens with biological information,providing a basis for theoretical research,which can effectively prevent allergies,or establish a rapid and sensitive detection method to detect sesame in food,which can play a monitoring role.Firstly,the whole protein in white sesame was extracted by salt extraction,and the allergenicity of sesame allergen was studied.It was found that boiling treatment could reduce the solubility and antigenicity of sesame protein.The detection rate of ELISA was70.67%.BCA kit The detection rate was 63.40%.High temperature and high pressure had no effect on it.Roasting and microwave can appropriately reduce the solubility and antigenicity of sesame protein.Since these treatments cannot effectively reduce the antigenicity of sesame allergens,food containing sesame protein after deep processing still has the risk of allergy,so further analysis of sesame allergens is required.The biological information of sesame allergen(SA)was then analyzed by computer.Analysis of homology between sesame allergens and other nut allergens.Then,the epitope prediction of sesame allergen was performed,and the three-dimensional structure of sesame protein was predicted by homology modeling.The results of the pull-type patterning showed that the reliability of the three-dimensional structure was good,which provided a reference for the use of bioinformatics methods to simulate the antigenic epitopes of food allergens.Finally,two immunization methods for rapid detection of sesame allergens were established.One was a colloidal gold immunocolorimetric assay based on magnetic separation.The optimal detection conditions of this method are: the optimal connection amount of magnetic spheres(MPMs)was 20 μg,the addition amount of gold-labeled antibody(Au NPs-Ab)was 6 μL,the optimal p H range was 5.5-7.4,the optimized ion when the concentration was less than 0.01 M,it will not affect the experiment.Under this optimal condition,the change of absorbance of the established standard curve and the concentration of sesame allergen showed a good linear trend in the range of 50-800 μg/L,and the limit of detection(LOD)was 45.529 μg/L.A second fluoroimmunoassay based on magnetic separation was subsequently established.After optimization,the best working conditions were as follows: the optimal connection amount of the magnetic sphere was 20μg,the optimal addition amount of the signal probe(QDs-Ab)was 60 μL,the p H detection range of the sample was 5.5-8.5,the optimized ion When the concentration was controlled below 0.01 M,QDs will not decay and will not affect the binding of antigen and antibody.Under this optimal condition,the change of fluorescence intensity of the system and the concentration of sesame showed a good linear trend in the range of 80-640 μg/L,and the limit of detection(LOD)was 75.426 μg/L.