Method for detecting allergen in high oleic acid peanuts by mass spectrometry

Abstract

The invention provides a method for detecting an allergen in high oleic acid peanuts by using mass spectrometry. The method comprises the following steps: performing mass spectrometry on a high oleic acid peanut extract by using a mass spectrometry parallel reaction monitoring method to obtain protein information; comparing and screening the obtained protein information with bioinformatics data in a UniProt database to obtain the allergenic protein related to peanut allergy; and screening out a specific peptide fragment in the sensitization protein through a screening condition. The screening conditions comprise: 1) no modification; 2) the number of spectrums matching the amino acid sequence in the spectrum database is greater than 3; 3) the sequence only exists in one or two proteins; 4) the number of amino acids is 7 to 20; and 5) the peptide fragment does not contain cysteine and methionine. The method can be used for qualitatively and quantitatively detecting the peanut allergenic protein and is relatively high in accuracy; the protein and the peptide fragment can be confirmed, the corresponding peptide fragment can be specifically screened out, and a foundation is laid for subsequently detecting the peanut allergen by using an isotope dilution method.

Description

technical field [0001] The invention belongs to the technical field of allergen detection, and in particular relates to a method for detecting allergens in high-oleic peanuts by mass spectrometry. Background technique [0002] High oleic peanuts are peanut varieties with an oleic acid content of more than 70%. High oleic peanuts increase the content of oleic acid, reduce the content of linoleic acid, increase the ratio of oleic acid to linoleic acid, strengthen the anti-oxidation ability of peanuts, and improve storage stability. At present, there are insufficient studies on the allergenicity of high oleic acid peanuts. There are currently two types of methods commonly used to detect peanut allergens: one is based on the detection method of allergen component genes, such as: polymerase chain reaction (PCR method), real-time fluorescent polymerase chain reaction (RT-PCR), etc. The other is a detection method based on the allergenic protein, such as enzyme-linked immunosorbe...

AI Technical Summary

Problems solved by technology

However, since the heating process will change the primary structure of the allergenic protein, the antibody cannot correctly recognize the detected allergen, resulting in a false negative phenomenon

In addition, food components are complex, and processing will cause protein denaturation, and ELISA, an antigen-antibody detection mode, will also undergo cross-reaction, resulting in high detection values ​​and false positives, which will affect the accuracy of experimental results.

[0005] At present, Western blotting (WB) method is very mature. Western blotting has many advantages in detecting peanut allergenic proteins, but due to the experimental operation, it is easy to appear multiple bands, no bands, black spots in the background, gel Uneven staining and other phenomena affect the accuracy of the test. Therefore, Western blotting is mostly used for qualitative analysis of allergenic proteins

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