Techniques for identifying allergens and quantifying allergenicity

A number of techniques have been used to identify allergenic proteins, most being based on the principle of:

• Solubilising/extracting proteins

• Isolation of protein fractions

• Determining IgE binding ability of each fraction

• Characterisation of the protein/glycoprotein and larger-scale purification.

The techniques most often used in the current literature for allergen identification are:

• Separation on a gel such as SDS PAGE followed by Western blotting and immuno-labelling

• Separation by chromatography (often High Performance Liquid Chromatography) followed by ELISA.

Using ELISA or Western blotting, quantitative or semi-quantitative data on the binding of serum IgE to specific proteins can be calculated for individual patients. Generalisations on allergenicity of specific proteins in a food are made by assessing the proportion of affected individuals that have elevated IgE to that protein. These methods cannot predict the degree of symptoms that may be produced on exposure to each individual protein or the outcome of introducing novel foods into a community.

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