Method for determining key amino acid of milk allergen epitope

Abstract

The invention discloses a method for determining key amino acid of milk allergen epitope, which comprises the following steps of: synthesizing an epitope peptide section according to the primarily determined mimic epitopes area amino acid sequence, and fixing and protecting the epitope peptide section, wherein the C end covalence of the epitope peptide section is combined to an activated fibrous membrane and the N end is acetylized; taking single glycine as a negative control, reacting the synthesized amino acid sequence with polyclonal serum, and determining the main epitope of beta-milk globulin; sequentially substituting each amino acid with alanine or glycine to obtain the amino acid sequence needing to be synthesized; using the obtained amino acid sequence to synthesize the epitope peptide section, and fixing and protecting the epitope peptide section, wherein the C end covalence of the epitope peptide section is combined to the activated fibrous membrane and the N end is acetylized; and taking the single glycine as the negative control, reacting the synthesized amino acid sequence with the polyclonal serum, and determining the key amino acid in milk epitope. The method lays a foundation for the diagnosis and treatment of milk allergy and the detection of an allergen in foods and provides theoretical basis and technical support for the development of hypoallergenic milk.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the precise positioning of epitopes. Background technique [0002] Milk allergy is a common food allergy that affects 1-2% of adults and up to 8% of children under the age of three. There are nearly 20 proteins in cow's milk that are potentially allergenic. It has been reported that people with cow's milk allergies are allergic to cow's milk, which generally manifests as co-allergies to multiple proteins. β-lactoglobulin, α-lactalbumin, and casein are considered to be the main allergens, and other proteins such as bovine serum albumin, immunoglobulin, and lactoferrin also play a very important role in allergic reactions, with about 30% to 50% of milk allergies are allergic to these trace proteins, and some of them are only allergic to these trace proteins. [0003] Beta-lactoglobulin accounts for 10% of the total protein of milk and 50% of whey protein. β-lactoglobulin...

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Problems solved by technology

[0005] "Preliminary study on the localization of β-lactoglobulin allergen epitope in buffalo milk" ("Food Science" 2007, vol1.28, No.10, P360-363) discloses the β-lactoglobulin allergen in buffalo milk Preliminary results of epitope localization, that is, using immunoaffinity-purified specific rabbit serum to perform affinity panning on a phage random heptapeptide library, and to determine the epitope region of β-lactoglobulin allergen by using the network tool MIMOX analysis , but the key amino acids cannot be identified

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